SYNTHETIC AND RECOMBINANTLY PRODUCED COLLAGEN PEPTIDES WITH BIOLOGICAL EFFICACY

Description

[0001] The present invention relates to a synthetically or recombinantly produced collagen peptide for use in a method for the therapeutic treatment of the human or animal body having a molecular weight in a range of 0.18 to 5.0 kDa, in particular 1.1 to 5.0 kDa, as well as products comprising a collagen peptide according to the invention.

[0002] Collagen is an extracellular structural protein found in animals, such as mammals, birds, and fish. It is typically found in connective tissue, particularly as a component of the extracellular matrix. Tendons, ligaments, cartilage, and bones are particularly rich in collagen. However, collagens are not found in plants or single-celled organisms.

[0003] Collagens occur in various structurally and functionally distinct types and differ, among other things, in terms of their structure, function, and origin. The polypeptide chains that make up collagen are synthesized individually in the cell at the ribosomes of the endoplasmic reticulum in the form of larger precursor molecules and feature extensive repetitive (Gly-XY) n sequences, where X and Y can be any amino acid, but are usually proline and 4-hydroxyproline.

[0004] These precursor polypeptide chains are posttranslationally hydroxylated in the endoplasmic reticulum at proline and lysine residues of the polypeptide chain, forming hydroxyproline and hydroxylysine residues. Hydroxylation serves to stabilize neighboring collagen polypeptide chains in the right-handed triple helix (procollagen) that forms in the cell, consisting of three precursor polypeptide chains each.

[0005] The resulting procollagen is glycosylated intracellularly, secreted from the cell in the glycosylated triple-helical form (tropocollagen), and subsequently formed into collagen by peptidase-mediated cleavage of the terminal residues. This collagen assembles into collagen fibrils during a fibrillogenesis process, which are subsequently covalently cross-linked to form collagen fibers.

[0006] Collagen is also often used in denatured form, then referred to as gelatin, or in the form of hydrolysates.

[0007] If gelatin or collagen is subjected to hydrolytic processes, particularly enzymatic hydrolysis, to obtain collagen peptides, collagen hydrolysates with a wide range of compositions and application profiles can be produced depending on the collagen type and origin used, as well as the enzymatic conditions. However, these collagen hydrolysates represent a mixture of peptides whose molecular weights are distributed over specific size ranges. The use of such collagen hydrolysates, for example, as dietary supplements or as cosmetic aids, has long been known, among other things, for the prevention and / or treatment of conditions related to bones, joints, or connective tissue.

[0008] For example, WO 2012 / 065782 describes collagen hydrolysates obtained from pork rind gelatin, which serve to stimulate the biosynthesis of extracellular matrix proteins by skin cells and are particularly suitable for cosmetic purposes.

[0009] WO 2012 / 117012 discloses enzymatically hydrolyzed collagen from bovine split skin with an average molecular weight of 1500 to 8000 Da, which can be used together with a prebiotic for the prevention and / or treatment of osteoporosis.

[0010] WO 2018 / 041684 discloses the use of collagen hydrolysate to improve endurance performance and stimulate fat loss.

[0011] Although the use of collagen hydrolysates derived from animal materials offers advantages for many applications and consumer groups, the use of collagen hydrolysates derived from such materials may also be less desirable for certain consumer groups and application profiles. Certain consumer groups are fundamentally critical or opposed to raw materials derived from animal materials, whether due to concerns about contamination with harmful microorganisms or agents, such as processing aids, or undesirable immune reactions, or for religious or ethical reasons. Furthermore, the manufacturing processes used to obtain collagen hydrolysates derived from animal materials often involve complex and expensive digestion, purification, and further processing steps.

[0012] Against this background, it is not surprising that processes have been developed to produce gelatin, collagen, collagen hydrolysates and individual collagen peptides by biotechnological means using recombinant genetic engineering.

[0013] WO 2006 / 052451 A2 discloses the production of recombinant collagen type III in Pichia pastoris strains that also express human prolyl hydroxylases.

[0014] WO 2005 / 012356 A2 discloses the production of gelatin from human collagen type I and individual 50 kDa, 65 kDa and 100 kDa collagen peptide species, each in fully hydroxylated, partially and non-hydroxylated form.

[0015] Olsen et al. (Protein Expression and Purification, 2005, 40, pg. 346-357) discloses the recombinant production of an 8.5 kDa collagen peptide species from the α1 chain of human collagen in Pichia pastoris.

[0016] WO 01 / 34646 A2 also discloses the production of individual recombinant gelatin species with a defined molecular weight of 0 to 350 kDa resulting from the recombinant production route, which can be present in non-hydroxylated, partially or fully hydroxylated form.

[0017] The publication discloses the general applicability of individual recombinant gelatin species or mixtures thereof in the food, beverage, cosmetic, or pharmaceutical industries. The publication discloses further possible uses of the disclosed recombinant products as photographic compositions or as technical aids, e.g., in the semiconductor industry, paper production, or similar.

[0018] It is therefore known that recombinant gelatin or collagen peptides of any size, individually or in mixtures, hydroxylated or non-hydroxylated, can be used in a wide variety of technical or non-technical areas, for example in the food industry or in the therapy of human or animal diseases.

[0019] However, even in view of the increased demands of large sections of the population with regard to health, attractive appearance, mobility and fitness, even in advanced age, there is still a great need for food and preparations for the improvement and / or maintenance of health as well as for the treatment of diseases.

[0020] Health, mobility, physical attractiveness, and physical fitness are often closely linked to the condition of connective tissue. Connective tissue is a tissue found in all tissues of the body, for example, muscles, intestines, blood vessels, periodontal ligament, tendons, ligaments, bones, and skin, where it often performs supporting functions. Connective tissue typically has comparatively few cells and a high amount of intercellular substance, which usually contains extracellular matrix proteins, especially collagen.Connective tissues can occur in particular in the form of loose connective tissue, tight connective tissue, for example in the skin, tendons, ligaments and the meninges, gelatinous connective tissue and various other types of connective tissue, in particular supporting tissues, namely cartilage tissue and bone tissue, and they are particularly crucial for the mobility, external appearance, the integrity of the body's internal structures and the fitness of a body.

[0021] The technical problem underlying the present invention is therefore to provide agents and methods which have a biological effectiveness with regard to connective tissue, in particular connective tissue of the skin, tendons, ligaments, meninges, cartilage and bones, and thus make it possible, on the one hand, to treat diseases associated with this connective tissue and, on the other hand, to maintain a condition of such connective tissue which is considered desirable.

[0022] The present invention is also based on the technical problem of providing synthetic or recombinant collagen peptides which overcome the aforementioned disadvantages, in particular which can be produced recombinantly in a standardized, reliable and precisely defined form, even on a larger industrial and cost-effective scale, and which, due to their biological efficacy, are suitable for use in a method for the therapeutic treatment of the human or animal body.

[0023] The present invention solves the technical problem underlying it by providing the teachings of the independent claims, in particular also the teachings of the preferred embodiments in the description and the dependent claims.

[0024] The present invention relates to a synthetic or recombinant collagen peptide for use in a method for the prevention and / or treatment of bone diseases, cartilage diseases, degenerative joint diseases, diseases of the tendons or ligaments and / or treatment of skin diseases, wherein the collagen peptide has a molecular weight in a range of 0.18 to 5.0 kDa and is capable of stimulating the synthesis of extracellular matrix proteins, in particular collagen, in connective tissue cells, in particular in osteoblasts, chondrocytes and / or fibroblasts, and wherein the collagen peptide has an amino acid sequence selected from the group consisting of SEQ ID No. 1 to 30.

[0025] The synthetic or recombinant collagen peptides claimed according to the invention, in particular synthetically or recombinantly produced collagen peptides, with a molecular weight in a range of 0.18 to 5.0 kDa, in particular 1.1 to 5.0 kDa, advantageously exhibit biological activity, in particular in connective tissue of tendons and ligaments, in particular cartilage tissue and bone tissue, and are therefore particularly suitable for use in a method for the therapeutic or non-therapeutic treatment of the human or animal body. According to a preferred embodiment of the present invention, the synthetic or recombinant collagen peptide, in particular the synthetically or recombinantly produced collagen peptide, has the amino acid sequence SEQ ID No. 1, SEQ ID No. 6; preferably, the collagen peptide according to the invention consists of one of these amino acid sequences.

[0026] Preferably, the synthetic or recombinant collagen peptide, in particular the synthetically or recombinantly produced collagen peptide, has the amino acid sequence SEQ ID No. 1, preferably the collagen peptide according to the invention consists of this amino acid sequence.

[0027] Preferably, the synthetic or recombinant collagen peptide, in particular the synthetically or recombinantly produced collagen peptide, has the amino acid sequence SEQ ID No. 6, preferably the collagen peptide according to the invention consists of this amino acid sequence.

[0028] In a preferred embodiment of the present invention, the synthetic or recombinant collagen peptide, in particular the synthetically or recombinantly produced collagen peptide, has an amino acid sequence selected from SEQ ID Nos. 1 to 5, in particular selected from SEQ ID Nos. 1 to 4. The collagen peptide according to the invention preferably consists of one of these amino acid sequences. According to this embodiment, the synthetic or recombinant collagen peptide, in particular the synthetically or recombinantly produced collagen peptide, accordingly does not contain hydroxyproline in its amino acid sequence.

[0029] In a preferred embodiment of the present invention, the synthetic or recombinant collagen peptide, in particular the synthetically or recombinantly produced collagen peptide, has an amino acid sequence selected from SEQ ID Nos. 1 to 5, in particular selected from SEQ ID Nos. 1 to 4. The collagen peptide according to the invention preferably consists of one of these amino acid sequences, wherein at least one prolyl and / or at least one lysyl residue is hydroxylated. According to this embodiment, the synthetic or recombinant collagen peptide, in particular the synthetically or recombinantly produced collagen peptide, accordingly has at least one hydroxyproline and / or at least one hydroxylysine in its amino acid sequence.

[0030] In a further preferred embodiment of the present invention, the synthetic or recombinant collagen peptide, in particular the synthetically or recombinantly produced collagen peptide, has an amino acid sequence selected from SEQ ID Nos. 6 to 10, in particular selected from SEQ ID Nos. 6 to 9, preferably the collagen peptide according to the invention consists of one of these amino acid sequences.

[0031] According to a further preferred embodiment, the synthetic or recombinant collagen peptide, in particular the synthetically or recombinantly produced collagen peptide, has an amino acid sequence selected from the group consisting of SEQ ID Nos. 1 to 10, in particular selected from the group consisting of SEQ ID Nos. 1 to 4 and 6 to 9.

[0032] Preferably, the synthetic or recombinant collagen peptide, in particular the synthetically or recombinantly produced collagen peptide, consists of one of the amino acid sequences selected from the group consisting of SEQ ID Nos. 1 to 10, in particular selected from the group consisting of SEQ ID Nos. 1 to 4 and 6 to 9.

[0033] According to a further preferred embodiment of the present invention, the synthetic or recombinant collagen peptide, in particular the synthetically or recombinantly produced collagen peptide, has the amino acid sequence SEQ ID No. 15, SEQ ID No. 21, preferably the collagen peptide according to the invention consists of one of these amino acid sequences.

[0034] Preferably, the synthetic or recombinant collagen peptide, in particular the synthetically or recombinantly produced collagen peptide, has the amino acid sequence SEQ ID No. 15, preferably the collagen peptide according to the invention consists of this amino acid sequence.

[0035] Preferably, the synthetic or recombinant collagen peptide, in particular the synthetically or recombinantly produced collagen peptide, has the amino acid sequence SEQ ID No. 21, preferably the collagen peptide according to the invention consists of this amino acid sequence.

[0036] In a preferred embodiment of the present invention, the synthetic or recombinant collagen peptide, in particular the synthetically or recombinantly produced collagen peptide, has an amino acid sequence selected from SEQ ID No. 15 and SEQ ID No. 16. Preferably, the collagen peptide according to the invention consists of one of these amino acid sequences. According to this embodiment, the synthetic or recombinant collagen peptide, in particular the synthetically or recombinantly produced collagen peptide, therefore does not have hydroxyproline in its amino acid sequence.

[0037] In a preferred embodiment of the present invention, the synthetic or recombinant collagen peptide, in particular the synthetically or recombinantly produced collagen peptide, has an amino acid sequence selected from SEQ ID No. 15 and SEQ ID No. 16. The collagen peptide according to the invention preferably consists of one of these amino acid sequences, wherein at least one prolyl and / or at least one lysyl residue is hydroxylated. According to this embodiment, the synthetic or recombinant collagen peptide, in particular the synthetically or recombinantly produced collagen peptide, accordingly has at least one hydroxyproline and / or at least one hydroxylysine in its amino acid sequence.

[0038] In a further preferred embodiment of the present invention, the synthetic or recombinant collagen peptide, in particular the synthetically or recombinantly produced collagen peptide, has an amino acid sequence selected from SEQ ID Nos. 21 and 22, preferably the collagen peptide according to the invention consists of one of these amino acid sequences.

[0039] Particularly preferably, the synthetic or recombinant collagen peptide, in particular the synthetically or recombinantly produced collagen peptide, has the amino acid sequence SEQ ID No. 1, SEQ ID No. 15, preferably the collagen peptide according to the invention consists of one of these amino acid sequences.

[0040] According to a further preferred embodiment, the synthetic or recombinant collagen peptide, in particular the synthetically or recombinantly produced collagen peptide, has an amino acid sequence selected from the group consisting of SEQ ID Nos. 1 to 4, 6 to 9, 12 to 14 and 18 to 20.

[0041] In a further preferred embodiment of the present invention, the synthetic or recombinant collagen peptide, in particular the synthetically or recombinantly produced collagen peptide, has an amino acid sequence according to SEQ ID No. 7.

[0042] Preferably, the synthetic or recombinant collagen peptide, in particular the synthetically or recombinantly produced collagen peptide, consists of the amino acid sequence according to SEQ ID No. 7.

[0043] In a further preferred embodiment of the present invention, the synthetic or recombinant collagen peptide, in particular the synthetically or recombinantly produced collagen peptide, has an amino acid sequence selected from SEQ ID Nos. 17 to 19, 21 and 22.

[0044] Preferably, the synthetic or recombinant collagen peptide, in particular the synthetically or recombinantly produced collagen peptide, consists of the amino acid sequence selected from SEQ ID Nos. 17 to 19, 21 and 22.

[0045] In a further preferred embodiment of the present invention, the synthetic or recombinant collagen peptide, in particular the synthetically or recombinantly produced collagen peptide, has an amino acid sequence selected from SEQ ID Nos. 27 to 30.

[0046] Preferably, the synthetic or recombinant collagen peptide, in particular the synthetically or recombinantly produced collagen peptide, consists of the amino acid sequence selected from SEQ ID Nos. 27 to 30.

[0047] In a particularly preferred embodiment, the synthetic or recombinant collagen peptide, in particular the synthetically or recombinantly produced collagen peptide, according to the present invention is a non-hydroxylated collagen peptide.

[0048] In a further preferred embodiment, the synthetic or recombinant collagen peptide, in particular the synthetically or recombinantly produced collagen peptide, according to the present invention is a hydroxylated collagen peptide.

[0049] Preferably, the synthetic or recombinant collagen peptide, in particular the synthetically or recombinantly produced collagen peptide, comprises hydroxylated proline and / or hydroxylated lysine.

[0050] Preferably, the synthetic or recombinant collagen peptide, in particular the synthetically or recombinantly produced collagen peptide, is a non-hydroxylated, partially hydroxylated, or fully hydroxylated collagen peptide. According to another particularly preferred embodiment, any proline present in the synthetic or recombinant collagen peptide is hydroxylated.

[0051] According to a preferred embodiment of the present invention, the synthetic or recombinant collagen peptide, in particular the synthetically or recombinantly produced collagen peptide, is glycosylated. Preferably, the synthetic or recombinant collagen peptide, in particular the synthetically or recombinantly produced collagen peptide, is glycosylated at at least one hydroxylated lysine. Preferably, each hydroxylated lysine of the synthetic or recombinant collagen peptide, in particular the synthetically or recombinantly produced collagen peptide, is glycosylated.

[0052] In a preferred embodiment of the present invention, the collagen peptide according to the invention has an amino acid sequence occurring in collagen of types I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV, XVI, XVII, XVIII, XIX, XX, XXI, XXII, XXIII, XXIV, XXV, XXVI, XXVII, preferably type I, II or III, preferably type I, preferably type II, preferably type III. The collagen peptide according to the invention preferably has an amino acid sequence occurring in collagen from vertebrates, in particular from fish, amphibians, reptiles, birds and mammals, in particular in human, bovine, porcine, equine or avian collagen of types I, II or III, preferably type I, preferably type II, preferably type III.

[0053] Particularly preferably, the collagen peptide according to the invention has an amino acid sequence occurring in human collagen, in particular in human type I collagen, preferably in the α1 chain of human type I collagen.

[0054] Particularly preferably, the collagen peptide according to the invention has an amino acid sequence occurring in non-human collagen, in particular in non-human type I collagen, preferably in the α1 chain of the non-human type I collagen, in particular an amino acid sequence occurring in bovine, porcine, equine or avian collagen.

[0055] In a preferred embodiment of the present invention, the synthetic or recombinant collagen peptide, in particular the synthetically or recombinantly produced collagen peptide, is collagenase-resistant, in particular resistant to digestion by human collagenases.

[0056] According to a further preferred embodiment of the present invention, the synthetic or recombinant collagen peptide, in particular the synthetically or recombinantly produced collagen peptide, is capable of stimulating the synthesis of extracellular matrix proteins, such as collagen, proteoglycan and / or elastin, in particular collagen, in connective tissue cells, in particular in osteoblasts, chondrocytes and / or fibroblasts.

[0057] In a preferred embodiment of the present invention, the collagen peptide according to the invention is a synthetically produced collagen peptide, preferably a collagen peptide produced by chemical synthesis, in particular solid-phase synthesis, preferably Merrifield synthesis, Bailey peptide synthesis, or chemoenzymatic synthesis. In a further preferred embodiment of the present invention, the collagen peptide according to the invention is a recombinantly produced collagen peptide.

[0058] In a preferred embodiment of the present invention, the synthetic or recombinant collagen peptide, in particular the synthetically or recombinantly produced collagen peptide, has no amino acid modification, in particular no hydroxylation. Particularly preferably, the synthetic or recombinant collagen peptide, in particular the synthetically or recombinantly produced collagen peptide, has no hydroxylated and / or glycosylated amino acids.

[0059] According to a preferred embodiment of the present invention, the synthetic or recombinant collagen peptide, in particular the synthetically or recombinantly produced collagen peptide, for use in a method for the therapeutic treatment of the human or animal body has a molecular weight in a range of 0.18 to 5.0 kDa, preferably 0.2 to 5.0 kDa, preferably 0.3 to 5.0 kDa, preferably 0.4 to 5.0 kDa, preferably 0.5 to 5.0 kDa, preferably 0.6 to 5.0 kDa, preferably 0.7 to 5.0 kDa, preferably 0.8 to 5.0 kDa, preferably 0.9 to 5.0 kDa, preferably 1.0 to 5.0 kDa, preferably 1.1 to 5.0 kDa, preferably 1.2 to 5.0 kDa, preferably 1.3 to 5.0 kDa, preferably 1.4 to 5.0 kDa, preferably 1.5 to 5.0 kDa, preferably 1.6 to 5.0 kDa, preferably 1.7 to 5.0 kDa, preferably 1.8 to 5.0 kDa, preferably 1.9 to 5.0 kDa, preferably 2.0 to 5.0 kDa, preferably 2.1 to 4.9 kDa, preferably 2.2 to 4.8 kDa, preferably 2.3 to 4.7 kDa, preferably 2.4 to 4.6 kDa, preferably 2,5 to 4.5 kDa, preferably 1.2 to 3.2, preferably 1.3 to 3.0 kDa, preferably 1.5 to 3.0 kDa, preferably 1.8 to 3.0 kDa, preferably 2.0 to 3.0 kDa.

[0060] The present invention also relates to the synthetic or recombinant collagen peptide according to the invention, in particular the synthetically or recombinantly produced collagen peptide, for use in a method for the prevention and / or treatment of bone diseases, in particular osteoporosis.

[0061] Disclosed (not according to the invention) are synthetic or recombinant collagen peptides, in particular the synthetically or recombinantly produced collagen peptide, for use in a method for the prevention and / or treatment of sarcopenia.

[0062] Disclosed (not according to the invention) are synthetic or recombinant collagen peptides, in particular the synthetically or recombinantly produced collagen peptide, for use in a method for the prevention and / or treatment of degenerative loss of muscle mass.

[0063] In a preferred embodiment, the present invention relates to the synthetic or recombinant collagen peptide according to the invention, in particular the synthetically or recombinantly produced collagen peptide, for use in a method for the prevention and / or treatment of cartilage diseases, in particular osteoarthritis or arthritis.

[0064] Disclosed (not according to the invention) are synthetic or recombinant collagen peptides, in particular the synthetically or recombinantly produced collagen peptide, for use in a method for improving muscle strength.

[0065] Disclosed (not according to the invention) are synthetic or recombinant collagen peptides, in particular the synthetically or recombinantly produced collagen peptide, for use in a method for the prevention and / or treatment of a pathological condition characterized by reduced mitochondrial activity, in particular for the prevention and / or treatment of a pathological condition characterized by reduced endurance performance.

[0066] Disclosed (not according to the invention) are synthetic or recombinant collagen peptides, in particular the synthetically or recombinantly produced collagen peptide, for use in a method for stimulating fat loss.

[0067] Disclosed (not according to the invention) are synthetic or recombinant collagen peptides, in particular the synthetically or recombinantly produced collagen peptide, for use in a method for reducing body weight.

[0068] In a preferred embodiment, the present invention relates to the synthetic or recombinant collagen peptide according to the invention, in particular the synthetically or recombinantly produced collagen peptide, for use in a method for the prevention and / or treatment of osteoarthritis, rheumatoid arthritis, rheumatic diseases, spondylitis and / or fibromyalgia.

[0069] In a preferred embodiment, the present invention relates to the synthetic or recombinant collagen peptide according to the invention, in particular the synthetically or recombinantly produced collagen peptide, for use in a method for the prevention and / or treatment of psoriasis vulgaris, acne, atopic dermatitis, chronic pruritus and / or rosacea.

[0070] In a preferred embodiment, the present invention relates to the synthetic or recombinant collagen peptide according to the invention, in particular the synthetically or recombinantly produced collagen peptide, for use in a method for treating wounds, in particular chronic wounds, acute wounds and / or burns.

[0071] Disclosed (not according to the invention) are synthetic or recombinant collagen peptides, in particular the synthetically or recombinantly produced collagen peptide, for use in a method for the prevention and / or treatment of degenerative nerve diseases.

[0072] Disclosed (not according to the invention) are synthetic or recombinant collagen peptides, in particular the synthetically or recombinantly produced collagen peptide, for use in a method for the prevention and / or treatment of dementia.

[0073] Disclosed (not according to the invention) are synthetic or recombinant collagen peptides, in particular the synthetically or recombinantly produced collagen peptide, for use in a method for the prevention and / or treatment of Alzheimer's disease.

[0074] Disclosed (not according to the invention) are synthetic or recombinant collagen peptides, in particular the synthetically or recombinantly produced collagen peptide, for use in a method for the prevention and / or treatment of a pathological condition characterized by a reduction in mental performance.

[0075] Disclosed (not according to the invention) are synthetic or recombinant collagen peptides, in particular the synthetically or recombinantly produced collagen peptide, for use in a method for the prevention and / or treatment of diseases associated with dysfunctions of the blood-brain barrier, in particular the structure and / or function of the meninges.

[0076] Disclosed (not according to the invention) are synthetic or recombinant collagen peptides, in particular the synthetically or recombinantly produced collagen peptide, for use in a method for the prevention and / or treatment of intestinal diseases, in particular chronic inflammatory bowel diseases.

[0077] Disclosed (not according to the invention) are synthetic or recombinant collagen peptides, in particular the synthetically or recombinantly produced collagen peptide, for use in a method for the prevention and / or treatment of diseases of the cardiovascular system, in particular the structure and / or function of the blood vessels, in particular the vascular wall, in particular for the prevention and / or treatment of high blood pressure and / or circulatory disorders.

[0078] Disclosed (not according to the invention) are synthetic or recombinant collagen peptides, in particular the synthetically or recombinantly produced collagen peptide, for use in a method for the prevention and / or treatment of diseases of the periodontium.

[0079] The present invention further relates to a non-therapeutic method using the collagen peptide according to the invention for the optical improvement of the skin, in particular for reducing the formation of wrinkles, improving skin elasticity, increasing skin tone, increasing skin moisture content, reducing cellulite and / or reducing stretch marks, in particular stretch marks.

[0080] The present invention also relates to a non-therapeutic method using the collagen peptide according to the invention for accelerating nail growth and / or reducing nail brittleness.

[0081] The present invention also relates to a non-therapeutic method using the collagen peptide according to the invention for optical and structural improvement of the hair, in particular for improving hair quality, reducing split ends and / or reducing / delaying hair loss.

[0082] Disclosed (not according to the invention) is the non-therapeutic use of the collagen peptide according to the invention for increasing the number and / or activity of mitochondria.

[0083] Disclosed (not according to the invention) is the non-therapeutic use of the collagen peptide according to the invention for improving endurance performance.

[0084] Disclosed (not according to the invention) is the non-therapeutic use of the collagen peptide according to the invention for improving mental performance.

[0085] In a preferred embodiment of the present invention, the synthetic or recombinant collagen peptide according to the invention is used alone, i.e. without further substances, for use in one of the applications provided for by the invention.

[0086] In a further embodiment of the present invention, the synthetic or recombinant collagen peptide according to the invention is used as the only agent having biological activity in an application provided according to the invention.

[0087] In a further preferred embodiment, the synthetic or recombinant collagen peptide according to the invention is used together with at least one further agent, in particular a further biologically active agent, in an application provided according to the invention.

[0088] The present invention also relates to a pharmaceutical composition according to the claims, comprising a collagen peptide according to the invention and at least one pharmaceutically acceptable additive, as well as the pharmaceutical composition for use in a method for the therapeutic treatment of the human or animal body, in particular for use in at least one of the aforementioned indications. Accordingly, the collagen peptide according to the invention can be administered in the form of a pharmaceutical composition. The pharmaceutical composition according to the invention is particularly advantageously administered, for example, in the form of tablets, lozenges, chewable tablets, capsules, bite-sized capsules, coated tablets, pastilles, syrups, gels, or ointments.

[0089] The present invention further relates to a dietary supplement according to the claims, comprising a collagen peptide according to the invention and at least one food-acceptable additive, as well as the dietary supplement for use in a method for the therapeutic treatment of the human or animal body, in particular for use in at least one of the aforementioned indications. Accordingly, the collagen peptide according to the invention can be administered in the form of a dietary supplement. The dietary supplement according to the invention is particularly advantageously in the form of a tablet, coated tablet, pastille, sachet, solution, suspension, or gel, for example, in an ampoule, as granules, or powder. Due to its good solubility, the collagen peptide according to the invention can also be added to various beverages without causing clouding.

[0090] The present invention also relates to a cosmetic product according to the claims, comprising a collagen peptide according to the invention and at least one skin-compatible additive, as well as the cosmetic product for use in a method for the therapeutic treatment of the human or animal body, in particular for use in at least one of the aforementioned indications. Accordingly, the collagen peptide according to the invention can be administered in the form of a cosmetic composition. The cosmetic composition according to the invention is particularly advantageously administered, for example, in the form of lotions, ointments, creams, gels, powders, syringes, or sprays.

[0091] The invention also relates to a food or luxury item according to the claims, comprising a collagen peptide according to the invention, as well as the food or luxury item for use in a method for the therapeutic treatment of the human or animal body, in particular for use in at least one of the aforementioned indications. According to a preferred embodiment, the food or luxury item is a chocolate bar, protein bar, cereal bar, instant powder for preparing beverages, milk, dairy products, for example yogurt, whey or quark, and milk substitutes, for example soy milk, rice milk, almond milk and coconut milk (so-called functional food) or a beverage, e.g. a soft drink or fitness drink.

[0092] If the collagen peptide according to a preferred embodiment of the invention is not used as the sole physiologically active component of a product, in particular a pharmaceutical composition, a dietary supplement, a cosmetic composition or a food or beverage, it can be combined with one or more other components that have a positive effect on general health, in particular on endurance performance.Such components are preferably selected from the group consisting of vitamin C, vitamins of the B, D, E and K series, omega-3 fatty acids, omega-6 fatty acids, conjugated linolenic acids, caffeine and its derivatives, guarana extract, rosehip extract, green tea extract, epigallocatechin gallate, creatine, L-carnitine, α-lipoic acid, N-acetylcysteine, NADH, D-ribose, magnesium aspartate, antioxidants such as anthocyanins, carotenoids, flavonoids, resveratrol, glutathione and superoxide dismutase (SOD), cannabinoids such as cannabidiol (CBD), adaptogens such as. Rhodiola rosea, Panax ginseng, Withania somnifera, Shiitake, Ganoderma lucidum Lepidium meyenii, Minerals such as iron, magnesium, calcium, zinc, selenium and phosphorus, as well as other proteins, hydrolysates and peptides such as soy, wheat and whey protein.

[0093] In a further preferred embodiment of the present invention, the products according to the invention, in particular the pharmaceutical composition, the dietary supplement, the cosmetic composition or the food or luxury item, contain no further proteins or peptides, in particular no further collagen peptides, in addition to the collagen peptide according to the invention.

[0094] In a preferred embodiment of the invention, the collagen peptide is administered in an amount of 1 to 40 g per day, preferably from 1 to 30 g per day, preferably from 1 to 20 g per day, preferably from 1 to 15 g per day, preferably from 2.5 to 30 g per day, preferably 2.5 to 20 g per day, preferably 2.5 to 15 g per day, preferably 2.5 to 10 g per day, preferably 4 to 15 g per day, preferably 4 to 12 g per day, more preferably from 5 to 25 g per day, preferably 5 to 15 g per day, preferably 10 to 25 g per day, preferably 12 to 22 g per day, preferably 6 to 15 g per day, in particular from 2.5 to 7.5 g per day, preferably 2.5 to 5 g per day.

[0095] The present invention also relates to methods for the prevention and / or treatment of the aforementioned indications, in particular the aforementioned therapeutic indications, according to which an amount of at least one of the recombinant or synthetically produced peptides according to the invention, optionally with an additive, which is sufficient for the therapeutic purpose is administered to the human or animal body.

[0096] Also disclosed (not according to the invention) is a non-therapeutic method for improving muscle strength, increasing muscle mass, stimulating fat loss, reducing body weight, maintaining and / or improving bone health, maintaining and / or improving skin health, maintaining and / or improving intestinal health, maintaining and / or improving blood vessel structure, maintaining and / or improving the health of the cardiovascular system, maintaining and / or improving the periodontal ligament, maintaining and / or improving the health of the nails and hair of a human or animal body, maintaining and / or increasing the number and / or activity of mitochondria, maintaining and / or improving endurance performance or maintaining and / or improving mental performance,wherein at least one collagen peptide according to the invention is administered to the human or animal body.

[0097] Also disclosed (not according to the invention) is a process for producing a collagen peptide according to the invention, comprising the process steps: a) Providing an expression system comprising at least one expression cassette, wherein the expression cassette comprises at least one nucleotide sequence encoding a collagen peptide having a molecular weight in a range of 0.18 to 10 kDa, in particular 0.18 to 5.0 kDa, in particular 1.1 to 5.0 kDa, b) Cultivating the expression system under conditions enabling the expression of the collagen peptide, and c) Obtaining the collagen peptide according to the invention.

[0098] The method for producing a collagen peptide according to the invention, in particular a synthetic or recombinant collagen peptide for the therapeutic treatment of the human or animal body with a molecular weight in a range from 0.18 to 10.0 kDa, in particular 0.18 to 5.0 kDa, in particular 1.1 to 5.0 kDa, is characterized in particular in that a precisely defined, recombinantly produced collagen peptide with a molecular weight in a range from 0.18 to 10.0 kDa, in particular 0.18 to 5.0 kDa, in particular 1.1 to 5.0 kDa, is obtained, which is suitable for use in a method for the therapeutic treatment of the human or animal body, in particular due to its biological effectiveness.

[0099] Due to its recombinant production method, the collagen peptide provided according to the invention exhibits a particularly high degree of purity compared to collagen peptides obtained hydrolytically from natural sources. It can be produced in a wide variety of expression systems, even on an industrial scale, without undesirable contamination. The collagen peptide according to the invention also advantageously exhibits biological activity, particularly with regard to applications in a method for the therapeutic treatment of the human or animal body.

[0100] The biological efficacy of the collagen peptides according to the invention, and thus their suitability for use in a method for the therapeutic treatment of the human or animal body, advantageously already applies to the collagen peptides obtained directly from the method according to the invention, without the need for further processing steps. Thus, both the hydroxylated and the non-hydroxylated collagen peptides according to the present invention exhibit biological efficacy, in particular at least the same biological efficacy as collagen peptides obtained from natural sources of the same molecular weight and / or mixtures of collagen peptides obtained from natural sources with a comparable average molecular weight.It is particularly advantageous that the collagen peptides according to the invention surprisingly have a biological activity even in non-hydroxylated form, preferably the same biological activity as hydroxylated collagen peptides obtained from natural sources of the same molecular weight and / or mixtures of hydroxylated collagen peptides obtained from natural sources with a comparable average molecular weight, particularly preferably a better biological activity than hydroxylated collagen peptides obtained from natural sources of the same molecular weight and / or mixtures of hydroxylated collagen peptides obtained from natural sources with a comparable average molecular weight.

[0101] Preferably, the hydroxylated as well as the non-hydroxylated collagen peptides according to the present invention show a biological activity in osteoblasts, fibroblasts and / or chondrocytes, preferably in osteoblasts, preferably in fibroblasts, preferably in chondrocytes, preferably in osteoblasts and fibroblasts, preferably in osteoblasts and chondrocytes, preferably in fibroblasts and chondrocytes, preferably in fibroblasts and chondrocytes, preferably at least the same biological activity as collagen peptides obtained from natural sources of the same molecular weight and / or mixtures of collagen peptides obtained from natural sources with a comparable average molecular weight, particularly preferably a better biological activity than collagen peptides obtained from natural sources of the same molecular weight and / or mixtures of collagen peptides obtained from natural sources with a comparable average molecular weight.

[0102] The biological activity found according to the invention and attributed to the synthetic or recombinant collagen peptides, in particular the synthetically or recombinantly produced collagen peptides of the present invention, in particular in connective tissue, in particular in connective tissue of the skin, intestine, blood vessels and periodontium, meninges, tendons, ligaments, cartilage and / or bones, can be determined in particular by means of the in vitro tests presented in Examples 3 to 6 for stimulating the synthesis of extracellular matrix proteins in osteoblasts, fibroblasts and chondrocytes.

[0103] In a preferred embodiment, the synthetic or recombinant collagen peptides according to the invention, in particular the synthetically or recombinantly produced collagen peptides, in particular the synthetically or recombinantly produced hydroxylated collagen peptides, in particular the synthetically or recombinantly produced non-hydroxylated collagen peptides, of the present invention have a biological activity, in particular they have a biological activity in at least one, preferably in at least two, preferably in all, of the in vitro tests presented in Examples 3 to 6 for stimulating the synthesis of extracellular matrix proteins in osteoblasts, fibroblasts and chondrocytes.

[0104] Preferably, the synthetic or recombinant collagen peptides according to the invention, in particular the synthetically or recombinantly produced collagen peptides, in particular the synthetically or recombinantly produced hydroxylated collagen peptides, in particular the synthetically or recombinantly produced non-hydroxylated collagen peptides, of the present invention have the same biological activity as collagen peptides of the same molecular weight isolated from natural sources, in particular they have the same biological activity as collagen peptides of the same molecular weight isolated from natural sources in at least one, preferably in at least two, preferably in all, of the in vitro tests presented in Examples 3 to 6 for stimulating the synthesis of extracellular matrix proteins in osteoblasts, fibroblasts and chondrocytes.

[0105] Particularly preferably, the synthetic or recombinant collagen peptides according to the invention, in particular the synthetically or recombinantly produced collagen peptides, in particular the synthetically or recombinantly produced hydroxylated collagen peptides, in particular the synthetically or recombinantly produced non-hydroxylated collagen peptides, of the present invention have the same biological activity as mixtures of collagen peptides isolated from natural sources, in particular as mixtures of collagen peptides isolated from natural sources with a comparable average molecular weight, in particular they have in at least one, preferably in at least two, preferably in all, of the in vitro tests presented in Examples 3 to 6 for stimulating the synthesis of extracellular matrix proteins in osteoblasts,Fibroblasts and chondrocytes have the same biological efficacy as mixtures of collagen peptides isolated from natural sources, in particular as mixtures of collagen peptides isolated from natural sources with comparable average molecular weight.

[0106] Particularly preferably, the synthetic or recombinant collagen peptides according to the invention, in particular the synthetically or recombinantly produced collagen peptides, in particular the synthetically or recombinantly produced hydroxylated collagen peptides, in particular the synthetically or recombinantly produced non-hydroxylated collagen peptides, of the present invention have a better biological activity than collagen peptides of the same molecular weight isolated from natural sources, in particular they have a better biological activity in at least one, preferably in at least two, preferably in all, of the in vitro tests presented in Examples 3 to 6 for stimulating the synthesis of extracellular matrix proteins in osteoblasts, fibroblasts and chondrocytes than collagen peptides of the same molecular weight isolated from natural sources.

[0107] Particularly preferably, the synthetic or recombinant collagen peptides according to the invention, in particular the synthetically or recombinantly produced collagen peptides, in particular the synthetically or recombinantly produced hydroxylated collagen peptides, in particular the synthetically or recombinantly produced non-hydroxylated collagen peptides, of the present invention have a better biological efficacy than mixtures of collagen peptides isolated from natural sources, in particular mixtures of collagen peptides isolated from natural sources with a comparable average molecular weight, in particular they have in at least one, preferably in at least two, preferably in all, of the in vitro tests presented in Examples 3 to 6 for stimulating the synthesis of extracellular matrix proteins in osteoblasts,Fibroblasts and chondrocytes show better biological efficacy to mixtures of collagen peptides isolated from natural sources, especially to mixtures of collagen peptides isolated from natural sources with comparable average molecular weight.

[0108] The expression system provided in step a) may be a host cell, in particular a prokaryotic or eukaryotic cell.

[0109] The expression system can be a host cell selected from the group consisting of bacterial cell, yeast cell, fungal cell, mammalian cell, insect cell and plant cell.

[0110] The expression system, in particular the host cell, may be a bacterial cell, in particular of the species Escherichia coli or Bacillus subtilis.

[0111] The expression system, in particular the host cell, may be a yeast cell, in particular of the species Saccharomyces cerevisiae, Pichia pastoris or Ogataea angusta (Hansenula polymorpha).

[0112] The expression system, in particular the host cell, may be a fungal cell, in particular of the species Aspergillus niger.

[0113] The expression system, in particular the host cell, may be a mammalian cell, in particular a CHO cell, a HeLa cell or a HEK293 cell.

[0114] The expression system, in particular the host cell, may be an insect cell, in particular an Sf-9, Sf-21 or Tn-5 cell.

[0115] The expression system, in particular the host cell, may be a plant cell, in particular a maize or tobacco cell.

[0116] The expression system provided in step a) may be a host cell capable of hydroxylating proline, lysine, or proline and lysine residues of the expressed collagen peptide. The expression system provided in step a) may be a host cell capable of hydroxylating proline, lysine, or proline and lysine residues of the expressed collagen peptide.

[0117] The expression system provided in step a) may be an expression system exhibiting prolyl hydroxylase and / or lysyl hydroxylase activity. The expression system provided in step a) may be a host cell exhibiting prolyl hydroxylase and / or lysyl hydroxylase activity.

[0118] The expression system provided in step a) can be a host cell having at least one expression cassette comprising a prolyl 4-hydroxylase-encoding polynucleotide sequence. The expression system provided in step a) can be a host cell having at least one expression cassette comprising a prolyl 4-hydroxylase-encoding polynucleotide sequence, so that an in vivo hydroxylated collagen peptide is obtained in process step c).

[0119] The expression system provided in step a) can be a host cell having at least one expression cassette comprising a lysyl hydroxylase-encoding polynucleotide sequence. The expression system provided in step a) can be a host cell having at least one expression cassette comprising a lysyl hydroxylase-encoding polynucleotide sequence, so that an in vivo hydroxylated collagen peptide is obtained in process step c).

[0120] The expression system provided in step a) can be a host cell having at least one expression cassette comprising a prolyl 4-hydroxylase-encoding polynucleotide sequence and at least one expression cassette comprising a lysyl hydroxylase-encoding polynucleotide sequence. The expression system provided in step a) can be a host cell having at least one expression cassette comprising a prolyl 4-hydroxylase-encoding polynucleotide sequence and at least one expression cassette comprising a lysyl hydroxylase-encoding polynucleotide sequence, so that an in vivo hydroxylated collagen peptide is obtained in process step c).

[0121] Also disclosed (not according to the invention) is a process for producing a collagen peptide according to the invention, in particular an in vivo hydroxylated collagen peptide, comprising the process steps a) Providing an expression system comprising at least one expression cassette, wherein the expression cassette comprises at least one nucleotide sequence encoding a collagen peptide having a molecular weight in a range from 0.18 to 10.0 kDa, in particular 0.18 to 5.0 kDa, in particular 1.1 to 5.0 kDa, and wherein the expression system is capable of hydroxylating proline, lysine or proline and lysine residues of the expressed collagen peptide, b) Cultivating the expression system under conditions that enable the expression and hydroxylation of the collagen peptide, c) Obtaining the collagen peptide according to the invention, in particular the in vivo hydroxylated collagen peptide.

[0122] Using the aforementioned method, it is thus possible to obtain an in vivo hydroxylated, recombinantly produced collagen peptide with a specific molecular weight in a range of 0.18 to 10.0 kDa, in particular 0.18 to 5.0 kDa, in particular 1.1 to 5.0 kDa, which, depending on the cell-based expression system used, is characterized by a specific pattern of post-translational modifications, in particular hydroxylations and glycosylations. In this way, it is advantageously possible, in particular, to obtain directly, i.e., without the need for subsequent modification, a collagen peptide with the desired biological efficacy, in particular a recombinantly produced collagen peptide for use in a method for the therapeutic treatment of the human or animal body.

[0123] In a preferred embodiment, the produced recombinant in vivo hydroxylated collagen peptide has a biological activity, in particular it has a biological activity in at least one, preferably in at least two, preferably in all, of the in vitro tests presented in Examples 3 to 6 for stimulating the synthesis of extracellular matrix proteins in osteoblasts, fibroblasts and chondrocytes.

[0124] Preferably, the produced recombinant in vivo hydroxylated collagen peptide has the same biological activity as collagen peptides of the same molecular weight isolated from natural sources, in particular it has the same biological activity as collagen peptides of the same molecular weight isolated from natural sources in at least one, preferably in at least two, preferably in all of the in vitro tests presented in Examples 3 to 6 for stimulating the synthesis of extracellular matrix proteins in osteoblasts, fibroblasts and chondrocytes.

[0125] Particularly preferably, the produced recombinant in vivo hydroxylated collagen peptide has the same biological activity as mixtures of collagen peptides isolated from natural sources, in particular as mixtures of collagen peptides isolated from natural sources with a comparable average molecular weight, in particular it has the same biological activity in at least one, preferably in at least two, preferably in all, of the in vitro tests presented in Examples 3 to 6 for stimulating the synthesis of extracellular matrix proteins in osteoblasts, fibroblasts and chondrocytes as mixtures of collagen peptides isolated from natural sources, in particular as mixtures of collagen peptides isolated from natural sources with a comparable average molecular weight.

[0126] Particularly preferably, the produced recombinant in vivo hydroxylated collagen peptide has a better biological activity than collagen peptides of the same molecular weight isolated from natural sources, in particular it has a better biological activity than collagen peptides of the same molecular weight isolated from natural sources in at least one, preferably in at least two, preferably in all, of the in vitro tests presented in Examples 3 to 6 for stimulating the synthesis of extracellular matrix proteins in osteoblasts, fibroblasts and chondrocytes.

[0127] Particularly preferably, the produced recombinant in vivo hydroxylated collagen peptide has a better biological activity than mixtures of collagen peptides isolated from natural sources, in particular mixtures of collagen peptides isolated from natural sources with a comparable average molecular weight, in particular it has a better biological activity in at least one, preferably in at least two, preferably in all, of the in vitro tests presented in Examples 3 to 6 for stimulating the synthesis of extracellular matrix proteins in osteoblasts, fibroblasts and chondrocytes than mixtures of collagen peptides isolated from natural sources, in particular mixtures of collagen peptides isolated from natural sources with a comparable average molecular weight.

[0128] The expression system provided in step a) may be an expression system which is not capable of causing hydroxylation of proline, lysine or proline and lysine residues of the expressed collagen peptide, in particular the expression system provided in step a) does not have prolyl hydroxylase and lysyl hydroxylase activity.

[0129] Also disclosed (not according to the invention) is a process for producing a collagen peptide according to the invention, in particular a non-hydroxylated collagen peptide, comprising the process steps a) Providing an expression system comprising at least one expression cassette, wherein the expression cassette comprises at least one nucleotide sequence encoding a collagen peptide having a molecular weight in a range from 0.18 to 10.0 kDa, in particular 0.18 to 5.0 kDa, in particular 1.1 to 5.0 kDa, and wherein the expression system is not capable of hydroxylating proline, lysine or proline and lysine residues of the expressed collagen peptide, b) Cultivating the expression system under conditions that enable the expression of the collagen peptide, c) Obtaining the collagen peptide according to the invention, in particular the non-hydroxylated collagen peptide.

[0130] In a preferred embodiment, the produced recombinant non-hydroxylated collagen peptide has a biological activity, in particular it has a biological activity in at least one, preferably in at least two, preferably in all, of the in vitro tests presented in Examples 3 to 6 for stimulating the synthesis of extracellular matrix proteins in osteoblasts, fibroblasts and chondrocytes.

[0131] Preferably, the recombinant non-hydroxylated collagen peptide produced has the same biological activity as collagen peptides of the same molecular weight isolated from natural sources, in particular it has the same biological activity as collagen peptides of the same molecular weight isolated from natural sources in at least one, preferably in at least two, preferably in all of the in vitro tests presented in Examples 3 to 6 for stimulating the synthesis of extracellular matrix proteins in osteoblasts, fibroblasts and chondrocytes.

[0132] Particularly preferably, the recombinant non-hydroxylated collagen peptide produced has the same biological activity as mixtures of collagen peptides isolated from natural sources, in particular as mixtures of collagen peptides isolated from natural sources with a comparable average molecular weight, in particular it has the same biological activity in at least one, preferably in at least two, preferably in all, of the in vitro tests presented in Examples 3 to 6 for stimulating the synthesis of extracellular matrix proteins in osteoblasts, fibroblasts and chondrocytes as mixtures of collagen peptides isolated from natural sources, in particular as mixtures of collagen peptides isolated from natural sources with a comparable average molecular weight.

[0133] Particularly preferably, the recombinant non-hydroxylated collagen peptide produced has a better biological activity than collagen peptides of the same molecular weight isolated from natural sources, in particular it has a better biological activity than collagen peptides of the same molecular weight isolated from natural sources in at least one, preferably in at least two, preferably in all, of the in vitro tests presented in Examples 3 to 6 for stimulating the synthesis of extracellular matrix proteins in osteoblasts, fibroblasts and chondrocytes.

[0134] Particularly preferably, the recombinant non-hydroxylated collagen peptide produced has a better biological activity than mixtures of collagen peptides isolated from natural sources, in particular mixtures of collagen peptides isolated from natural sources with a comparable average molecular weight, in particular it has a better biological activity in at least one, preferably in at least two, preferably in all, of the in vitro tests presented in Examples 3 to 6 for stimulating the synthesis of extracellular matrix proteins in osteoblasts, fibroblasts and chondrocytes than mixtures of collagen peptides isolated from natural sources, in particular mixtures of collagen peptides isolated from natural sources with a comparable average molecular weight.

[0135] According to a preferred embodiment of the present disclosure, the at least one nucleotide sequence of the at least one expression cassette is codon-optimized, i.e., those codons in the nucleotide sequence which are not or not preferentially used by the translation system of the provided expression system, in particular the provided cell-based expression system, in particular the provided host cell, are replaced by those which are preferentially used by the translation system of the provided expression system, in particular the provided cell-based expression system, in particular the provided host cell, without thereby changing the amino acid sequence of the encoded peptide or protein.

[0136] In a preferred embodiment of the present disclosure, the collagen peptide encoded by the nucleotide sequence is a collagen peptide of a vertebrate, in particular a mammal, for example a human or a non-human mammal, for example a horse, donkey, kangaroo, sheep, rodent, pig or cattle, a bird, for example a chicken, a fish, an amphibian, a reptile or an invertebrate, for example a jellyfish.

[0137] According to a preferred embodiment of the present invention, the at least one nucleotide sequence encodes a collagen peptide having a molecular weight in a range of 0.18 to 5.0 kDa, preferably 0.2 to 5.0 kDa, preferably 0.3 to 5.0 kDa, preferably 0.4 to 5.0 kDa, preferably 0.5 to 5.0 kDa, preferably 0.6 to 5.0 kDa, preferably 0.7 to 5.0 kDa, preferably 0.8 to 5.0 kDa, preferably 0.9 to 5.0 kDa, preferably 1.0 to 5.0 kDa, preferably 1.1 to 5.0 kDa, preferably 1.2 to 5.0 kDa, preferably 1.3 to 5.0 kDa, preferably 1.4 to 5.0 kDa, preferably 1.5 to 5.0 kDa, preferably 1.6 to 5.0 kDa, preferably 1.7 to 5.0 kDa, preferably 1.8 to 5.0 kDa, preferably 1.9 to 5.0 kDa, preferably 2.0 to 5.0 kDa, preferably 2.1 to 4.9 kDa, preferably 2.2 to 4.8 kDa, preferably 2.3 to 4.7 kDa, preferably 2.4 to 4.6 kDa, preferably 2.5 to 4.5 kDa, preferably 1.2 to 3.2, preferably 1.3 to 3.0 kDa, preferably 1.5 to 3.0 kDa, preferably 1.8 to 3.0 kDa, preferably 2.0 to 3.0 kDa.

[0138] In a preferred embodiment of the present invention, the collagen peptide produced by one of the aforementioned methods according to the set of claims is a non-hydroxylated, partially hydroxylated or fully hydroxylated collagen peptide, preferably a non-hydroxylated collagen peptide, preferably a partially hydroxylated collagen peptide, preferably a fully hydroxylated collagen peptide.

[0139] In a preferred embodiment of the present invention, the collagen peptide produced by one of the methods according to the claim set is a glycosylated collagen peptide. The collagen peptide is preferably glycosylated in vivo, preferably glycosylated ex vivo.

[0140] In a further preferred embodiment of the present invention, the collagen peptide produced by one of the methods is a non-glycosylated collagen peptide.

[0141] Also disclosed (not according to the invention) is a method for producing a collagen peptide according to the invention, wherein the at least one expression cassette of the expression system has several, preferably at least two, preferably at least three, preferably at least four, preferably at least five, consecutive nucleotide sequences, each encoding a collagen peptide having a molecular weight in a range of 0.18 to 10.0 kDa, in particular 0.18 to 5.0 kDa, in particular 1.1 to 5.0 kDa.

[0142] Thus, the expression system first synthesizes a precursor collagen peptide, from which collagen peptides according to the invention can subsequently be obtained by cleavage, in particular enzymatic cleavage.

[0143] The size of the precursor collagen peptide can be 2 to 100 kDa, preferably 3 to 80 kDa, preferably 4 to 60 kDa, preferably 5 to 50 kDa, preferably 10 to 45 kDa. The precursor collagen peptide can comprise 2 to 40 collagen peptides according to the invention, preferably 3 to 30 collagen peptides according to the invention, preferably 5 to 20 collagen peptides according to the invention.

[0144] The several, preferably at least two, preferably at least three, preferably at least four, preferably at least five, nucleotide sequences arranged in a row, each encoding a collagen peptide having a molecular weight in a range from 0.18 to 10.0 kDa, in particular 0.18 to 5.0 kDa, in particular 1.1 to 5.0 kDa, can be separated by sequences which allow cleavage, in particular enzymatic cleavage, of the collagen peptide expressed in step b) into several, at least two, preferably at least three, at least four, at least five, collagen peptides according to the invention.

[0145] Between steps b) and c), a step of cleaving the collagen peptide expressed in step b) can be carried out to obtain collagen peptides according to the invention, in particular collagen peptides with a molecular weight in a range from 0.18 to 10.0 kDa, in particular 0.18 to 5.0 kDa, in particular 1.1 to 5.0 kDa. Accordingly, in step b), a collagen peptide is expressed by the expression system provided in step a), which is then cleaved to obtain several, at least two, at least three, at least four, at least five, collagen peptides according to the invention.

[0146] The plurality of nucleotide sequences, preferably at least two, preferably at least three, preferably at least four, preferably at least five, arranged in series may be different nucleotide sequences. The plurality of nucleotide sequences, preferably at least two, preferably at least three, preferably at least four, preferably at least five, arranged in series may be identical nucleotide sequences.

[0147] The cleavage, in particular the enzymatic cleavage, of the collagen peptide expressed in step b), in particular the precursor collagen peptide, can be carried out by means of neutral, alkaline or acidic proteases.

[0148] The individual collagen peptides according to the invention of the expressed collagen peptide, in particular the precursor collagen peptide, can be separated from one another by specific recognition sequences.

[0149] The specific recognition sequences can be selected from the group consisting of factor Xa (Ile-(Glu / Asp)-Gly-Arg), TEV (Glu-Asn-Leu-Tyr-Phe-Gln-(Gly / Ser)), thrombin (Leu-Val-Pro-Arg-Gly-Ser), trypsin recognition sequence, papain recognition sequence.

[0150] In a preferred embodiment, according to the present invention, the collagen peptide is administered locally, in particular topically, or systemically, in particular enterally, preferably orally, or parenterally.

[0151] According to the invention, the term "biological activity" preferably refers to the ability of the collagen peptides according to the invention to stimulate the synthesis of extracellular matrix proteins, in particular the synthesis of collagen, proteoglycans and / or elastin, in cells, in particular osteoblasts, fibroblasts and / or chondrocytes.According to the invention, "biological efficacy" is preferably present when the incubation of cells, in particular osteoblasts, fibroblasts and / or chondrocytes, with the collagen peptides according to the invention results in a stimulation of the synthesis of extracellular matrix proteins, in particular the synthesis of collagen, proteoglycans and / or elastin, preferably measurable in at least one, preferably at least two, preferably in all of the in vitro tests presented in Examples 3 to 6 for stimulating the synthesis of extracellular matrix proteins in osteoblasts, fibroblasts and chondrocytes, compared to untreated cells and / or cells treated with a biologically inactive agent, in particular untreated osteoblasts, fibroblasts and / or chondrocytes, or osteoblasts, fibroblasts and / or chondrocytes treated with a biologically inactive agent.In a preferred embodiment of the present invention, a "biologically non-active agent" is understood to mean 160 Bloom gelatin from pig rinds or a 260 Bloom gelatin from beef split.

[0152] According to the invention, "equal biological activity" is preferably understood to mean that the collagen peptides according to the invention cause a stimulation of the synthesis of extracellular matrix proteins, in particular the synthesis of collagen, proteoglycans and / or elastin, in cells, in particular osteoblasts, fibroblasts and / or chondrocytes, which is comparable to the stimulation of cells, in particular osteoblasts, fibroblasts and / or chondrocytes, which, when incubated with a mixture of collagen peptides obtained from natural sources, in particular when incubated with a mixture of collagen peptides obtained from natural sources with a comparable average molecular weight, in particular with Verisol, produced according to EP 2640352 B1, Fortigel, produced according to WO 2010 / 149596 or Fortibone, produced according to WO 2014 / 072235 (EP 2916855 B1),can be measured. A "comparable stimulation" according to the invention preferably refers to a stimulation of the synthesis of extracellular matrix proteins, in particular the synthesis of collagen, proteoglycans and / or elastin, in cells, in particular osteoblasts, fibroblasts and / or chondrocytes, preferably measurable in at least one, preferably at least two, preferably in all, of the in vitro tests presented in Examples 3 to 6 for stimulating the synthesis of extracellular matrix proteins in osteoblasts, fibroblasts and chondrocytes, which deviates by at most 2%, preferably at most 1.5%, preferably at most 1% from the stimulation of the synthesis of extracellular matrix proteins, in particular the synthesis of collagen, proteoglycans and / or elastin, in cells, in particular osteoblasts, fibroblasts and / or chondrocytes, which is achieved by incubating the cells with a mixture of collagen peptides obtained from natural sources,in particular by incubating the cells with a mixture of collagen peptides obtained from natural sources with a comparable average molecular weight, in particular with Verisol, produced according to EP 2640352 B1, Fortigel, produced according to WO 2010 / 149596 or Fortibone, produced according to WO 2014 / 072235 (EP 2916855 B1).

[0153] The term "better biological efficacy" is preferably understood according to the invention to mean that the collagen peptides according to the invention cause a stronger stimulation of the synthesis of extracellular matrix proteins, in particular the synthesis of collagen, proteoglycans and / or elastin, in cells, in particular osteoblasts, fibroblasts and / or chondrocytes, than a mixture of collagen peptides obtained from natural sources, in particular a mixture of collagen peptides obtained from natural sources with a comparable average molecular weight, in particular with Verisol, produced according to EP 2640352 B1, Fortigel, produced according to WO 2010 / 149596 or Fortibone, produced according to WO 2014 / 072235 (EP 2916855 B1). According to the invention, "better biological efficacy" is preferably understood to mean an increase in the stimulation of the synthesis of extracellular matrix proteins, in particular the synthesis of collagen, proteoglycans and / or elastin, in cells,in particular osteoblasts, fibroblasts and / or chondrocytes, by the collagen peptides according to the invention by more than 2%, preferably at least 3%, preferably at least 4%, preferably at least 5%, compared to the stimulation of the synthesis of extracellular matrix proteins, in particular the synthesis of collagen, proteoglycans and / or elastin, in cells, in particular osteoblasts, fibroblasts and / or chondrocytes, by a mixture of collagen peptides obtained from natural sources, in particular with Verisol, produced according to EP 2640352 B1, Fortigel, produced according to WO 2010 / 149596 or Fortibone, produced according to WO 2014 / 072235 (EP 2916855 B1), wherein the increase in synthesis in at least one, preferably at least two, preferably in all, of the in vitro tests presented in Examples 3 to 6 for stimulating the synthesis of extracellular matrix proteins in osteoblasts, Fibroblasts and chondrocytes, is measurable.

[0154] A "biological activity", a "same biological activity" or a "better biological activity" can accordingly be present according to the invention if the collagen peptides according to the invention are capable of stimulating the synthesis of an extracellular matrix protein, preferably if at least the synthesis of one of the extracellular matrix proteins collagen, proteoglycan or elastin, preferably of two or three of these extracellular matrix proteins, is stimulated, in cells, preferably in at least one of the cell types osteoblasts, fibroblasts or chondrocytes, preferably in two or three of these cell types.Accordingly, in one embodiment of the present invention, biological efficacy can also be present if only the synthesis of one extracellular matrix protein is stimulated, in particular, for example, an extracellular matrix protein selected from the group consisting of collagen, proteoglycan, and elastin, while no stimulation of synthesis is observed for other extracellular matrix proteins in the cells in question. Likewise, biological efficacy, or an equal or better biological efficacy, can be present if stimulation of the synthesis of at least one extracellular matrix protein is observed in only one cell type, in particular a cell type selected from the group consisting of osteoblasts, fibroblasts, and chondrocytes, even if such stimulation does not occur in other cell types.It is preferably provided that the synthesis of more than one extracellular matrix protein, in particular of one or more of the explicitly mentioned extracellular matrix proteins selected from the group consisting of collagen, proteoglycan and elastin, is stimulated in cells, in particular in at least one of the cell types osteoblasts, fibroblasts or chondrocytes, in particular in two or three of these cell types.

[0155] According to the present invention, a "biological efficacy", a "same biological efficacy" or a "better biological efficacy" of the collagen peptides according to the invention in osteoblasts after incubation of the cells with the collagen peptides according to the invention can be determined by determining the expression of the mRNA of the corresponding extracellular matrix proteins, in particular selected from the group consisting of collagen, proteoglycan and elastin, in comparison to the expression of the mRNA of the corresponding extracellular matrix proteins, in particular selected from the group consisting of collagen, proteoglycan and elastin, in suitable controls by means of real-time PCR, in particular by the in vitro test listed in Example 3.

[0156] A "biological efficacy", a "same biological efficacy" or a "better biological efficacy" of the collagen peptides according to the invention in fibroblasts can be demonstrated according to the present invention after incubation of the cells with the collagen peptides according to the invention by determining the expression of the mRNA of the corresponding extracellular matrix proteins, in particular selected from the group consisting of collagen, biglycan and versican, in comparison to the expression of the mRNA of the corresponding extracellular matrix proteins, in particular selected from the group consisting of collagen, biglycan and versican, in suitable controls by means of real-time PCR, in particular by the in vitro test listed in Example 4.

[0157] According to the present invention, a "biological efficacy", an "equal biological efficacy" or a "better biological efficacy" of the collagen peptides according to the invention in chondrocytes can be determined after incubation of the cells with the collagen peptides according to the invention by radioactive labeling and detection of the amount of synthesized radioactively labeled collagen and / or by Alcian blue staining and photometric determination of the glycosaminoglycans (GAG) of synthesized proteoglycans, each in comparison to suitable controls, in particular by the in vitro tests listed in Example 5.

[0158] A "biological efficacy", a "same biological efficacy" or a "better biological efficacy" of the collagen peptides according to the invention in osteoblasts, fibroblasts and chondrocytes, in particular in fibroblasts and chondrocytes, can be determined according to the present invention after incubation of the cells with the collagen peptides according to the invention by determining the amount of synthesized extracellular matrix proteins, in particular selected from the group consisting of collagen, proteoglycan and elastin, particularly preferably selected from the group consisting of collagen and proteoglycan, in comparison to the determination of the amount of synthesized extracellular matrix proteins, in particular selected from the group consisting of collagen, proteoglycan and elastin, particularly preferably selected from the group consisting of collagen and proteoglycan,in suitable controls by photometric determination of the amount of dye-labelled synthesized extracellular matrix proteins, in particular selected from the group consisting of collagen, proteoglycan and elastin, particularly preferably selected from the group consisting of collagen and proteoglycan, in particular by the in vitro tests listed in Example 6.

[0159] According to the invention, the term "biological efficacy in at least one, preferably in at least two, preferably in all, of the in vitro tests presented in Examples 3 to 6 for stimulating the synthesis of extracellular matrix proteins in osteoblasts, fibroblasts, and chondrocytes" means that the biological efficacy of the collagen peptides according to the invention can be demonstrated using one of the assays explicitly described in Examples 3 to 6. The individual process parameters listed in Examples 3 to 6 can, if necessary, be varied according to the expert's understanding without affecting the fundamental significance of the test results.In a preferred embodiment of the present invention, the collagen peptides according to the invention exhibit biological efficacy in at least one, preferably in at least two, preferably in all, of the in vitro tests presented in Examples 3 to 6 for stimulating the synthesis of extracellular matrix proteins in osteoblasts, fibroblasts, and chondrocytes under exactly the process parameters listed in these examples. Furthermore, the biological efficacy of the collagen peptides according to the invention can also be demonstrated using other tests known to the person skilled in the art, in particular in vitro tests, preferably in vitro tests for stimulating the synthesis of extracellular matrix proteins in osteoblasts, fibroblasts, and chondrocytes.

[0160] According to the invention, the term "stimulation of the synthesis of extracellular matrix proteins" means stimulation of the biosynthesis of at least one protein of the extracellular matrix in cells, preferably in osteoblasts, fibroblasts and / or chondrocytes, and / or stimulation of the biosynthesis of at least one mRNA encoding a protein of the extracellular matrix in cells, preferably in osteoblasts, fibroblasts and / or chondrocytes, by exogenous influences, in particular by the collagen peptides according to the invention.In particular, the term "stimulation of the synthesis of extracellular matrix proteins" is understood according to the invention to mean an increase in the amount of at least one protein of the extracellular matrix secreted by cells, preferably osteoblasts, fibroblasts and / or chondrocytes, and / or an increase in the amount of at least one mRNA encoding a protein of the extracellular matrix synthesized in cells, preferably in osteoblasts, fibroblasts and / or chondrocytes, by exogenous influences, in particular by the collagen peptides according to the invention.

[0161] In the context of the present invention, the term "collagen" is understood in the art, in particular as defined, for example, in WO 01 / 34646. In a preferred embodiment, the term "collagen" refers to collagen types I to XXVII. In a further preferred embodiment, the term "collagen" is understood to mean a peptide having the sequence glycine-proline, glycine-4-hydroxyproline, or glycine-X-4-hydroxyproline, preferably the repetitive motif (Gly-XY) n , where X and Y can be any amino acid, preferably proline and 4-hydroxyproline. Particularly preferably, the term "collagen" is understood to mean a peptide having the repetitive motif (Gly-Pro-Y) n and / or (Gly-X-Hyp) m , where X and Y can be any amino acid.

[0162] The term "gelatin" is understood in the context of the present invention in the usual manner, in particular as defined, for example, in WO 01 / 34646.

[0163] In the context of the present invention, the term "collagen peptide" refers to a peptide that has an amino acid sequence found in collagen as defined above, wherein the peptide is at least a dipeptide, preferably an oligopeptide or polypeptide. The collagen peptide can be present in a chemically modified form, particularly hydroxylated and / or glycosylated form, or it can be unmodified.

[0164] In the context of the present disclosure, the term "amino acid modification" refers to a chemical modification of one or more amino acids before, after, or during the synthesis of the collagen peptide while retaining the original amino acid backbone, in particular one or more proteinogenic amino acids, of the collagen peptide. Thus, the term encompasses both the use of chemically modified amino acids for the synthesis of the collagen peptide according to the invention and the chemical modification of the amino acids after or during the synthesis of the collagen peptide. Amino acid modifications typical for collagen peptides include, in particular, hydroxylations of proline and lysine residues, as well as glycosylations of hydroxylated lysine residues. However, according to the invention, the term also encompasses other chemical modifications of amino acids, such as phosphorylations, N-glycosylations, acetylations, methylations, and myristoylations.

[0165] In the context of the present invention, a "synthetic or recombinant collagen peptide" or a "synthetically or recombinantly produced collagen peptide" is understood to mean a collagen peptide obtained by chemical synthesis, in particular solid-phase synthesis, or by biotechnological recombinant production using an expression system. According to the invention, the "synthetic collagen peptide" or "synthetically produced collagen peptide" and the "recombinant collagen peptide" or "recombinantly produced collagen peptide" have in common that they are not obtained from natural sources.

[0166] In the context of the present invention, the term "recombinant DNA" refers to an artificially produced or manipulated DNA molecule produced in vitro using genetic engineering methods. In a preferred embodiment, the recombinant DNA is composed of components from different organisms of origin.

[0167] In the context of the present invention, the term "expression cassette" is understood to mean a DNA segment which is responsible for the transcription of the information encoded in this segment into an RNA, in particular into an mRNA, and which has at least one promoter and one protein-coding nucleotide sequence, usually at least one promoter, at least one protein-coding nucleotide sequence and optionally a terminator.

[0168] In the context of the present invention, a "nucleotide sequence" is understood to mean the sequence of nucleotides of a nucleic acid, in particular a nucleic acid strand, in particular a DNA or RNA strand. A "nucleotide sequence" is therefore to be understood both as an informational unit and as the DNA or RNA strand that physically manifests this information.

[0169] In the context of the present invention, an "expression system" is understood to mean a system in which targeted and controlled protein biosynthesis can take place. According to the invention, the term "expression system" encompasses both cell-free expression systems, in which the components necessary for protein biosynthesis are not present within a cell, i.e., protein biosynthesis takes place outside a cell, and cell-based expression systems, in which protein biosynthesis takes place within a living cell. In the context of the present invention, a cell-free expression system is preferably a lysate or an extract from E. coli, Insect cells, wheat germ, tobacco cells, or mammalian cells, in particular CHO cells or rabbit reticulocytes, which contain the components necessary for protein biosynthesis, in particular a translation and transcription system. When using a cell-free expression system in one of the methods according to the present invention, the term "culturing" is synonymous with "incubating."

[0170] In the context of the present invention, a "host cell" is understood to mean a living cell capable of expressing peptides or proteins encoded in foreign DNA, in particular in recombinant DNA.

[0171] The term "obtaining the collagen peptide" according to process step c) refers to a process known to the person skilled in the art for isolating the collagen peptide from a composition containing several components by means of known isolation methods, such as centrifugation methods, in particular differential centrifugation and / or density gradient centrifugation, chromatographic methods, in particular gel filtration, ion exchange, affinity and / or high-performance liquid chromatography, electrophoresis methods, filtration methods and / or extraction methods, wherein an enrichment and purification of the component in question from the composition containing several components can preferably be achieved by sequential application of several isolation methods.

[0172] "Conditions enabling the expression of the collagen peptide" are understood to mean conditions such as, in particular, temperature, pressure, time, light, and the presence or absence of inducers and / or repressors that activate or enhance the expression of the collagen peptide. In a preferred embodiment, the expression of the collagen peptide takes place in a high-cell-density fermentation, in particular under high pressure, preferably high-pressure air. The specific conditions enabling the expression of the collagen peptide are known to the person skilled in the art and depend on the expression system and the expression cassette used, in particular the promoter contained therein. Depending on the structure of the expression cassette, the expression of the collagen peptide can be constitutive or inducible.

[0173] In the context of the present invention, the terms "comprising" and "having" are understood to mean that, in addition to the elements explicitly encompassed by these terms, further, not explicitly mentioned elements may be present. In the context of the present invention, these terms are also understood to mean that only the explicitly mentioned elements are encompassed and no further elements are present. In this particular embodiment, the meaning of the terms "comprising" and "having" is synonymous with the term "consisting of." Furthermore, the terms "comprising" and "having" also encompass compositions that, in addition to the explicitly mentioned elements, also contain further, not mentioned elements, which, however, are of a functionally and qualitatively subordinate nature. In this embodiment, the terms "comprising" and "having" are synonymous with the term "consisting essentially of."

[0174] If the first and second decimal places or the second decimal place are not specified in connection with the present invention, they are to be set as 0.

[0175] In the context of the present invention, the term "and / or" means that all members of a group linked by the term "and / or" are disclosed both alternatively to one another and cumulatively among themselves in any combination. For the expression "A, B and / or C," this means that the following disclosure content is to be understood: a) A or B or C or b) (A and B) or c) (A and C) or d) (B and C) or e) (A and B and C).

[0176] Further preferred embodiments emerge from the subclaims.

[0177] The invention is described below without limiting the general inventive concept using exemplary sequences, figures and embodiments.

[0178] This refers to: SEQ ID No. 1 the amino acid sequence Leu-Thr-Gly-Pro-Ile-Gly-Pro-Pro-Gly-Pro-Ala an 11 amino acid non-hydroxylated collagen peptide. SEQ ID No. 2 the amino acid sequence Gly-Ala-Pro-Gly-Lys-Asp-Gly-Val-Arg-Gly-Leu-Thr-Gly-Pro-Ile-Gly-Pro-Pro-Gly-Pro-Ala-Gly-Ala-Pro-Gly-Asp-Lys-Gly-Glu-Ala-Gly-Pro-Ser a 33 amino acid non-hydroxylated collagen peptide. SEQ ID No. 3 the amino acid sequence Gly-Ala-Pro-Gly-Lys-Asp-Gly-Val-Arg-Gly-Leu-Thr-Gly-Pro-Ile-Gly-Pro-Pro-Gly-Pro-Ala-Gly-Ala-Pro a 24 amino acid non-hydroxylated collagen peptide. SEQ ID No. 4 the amino acid sequence Gly-Ala-Pro-Gly-Lys-Asp-Gly-Val-Arg-Gly-Leu-Thr-Gly-Pro-Ile-Gly-Pro-Pro-Gly-Pro-Ala-Gly-Ala-Arg a 24 amino acid non-hydroxylated collagen peptide. SEQ ID No. 5 the amino acid sequence Gly-Ala-Pro-Gly-Pro-Pro-Gly-Pro-Pro-Gly-Ala-Arg-Gly-Gln-Ala-Gly-Val-Met-Gly-Phe-Pro-Gly-Pro-Lys a 24 amino acid non-hydroxylated collagen peptide. SEQ ID No. 6 the amino acid sequence Leu-Thr-Gly-Pro-Ile-Gly-Pro-4Hyp-Gly-Pro-Ala an 11 amino acid hydroxylated collagen peptide. SEQ ID No. 7 the amino acid sequence Gly-Ala-4Hyp-Gly-Lys-Asp-Gly-Val-Arg-Gly-Leu-Thr-Gly-Pro-Ile-Gly-Pro-4Hyp-Gly-Pro-Ala-Gly-Ala-4Hyp-Gly-Asp-Lys-Gly-Glu-Ala-Gly-Pro-Ser a 33 amino acid hydroxylated collagen peptide. SEQ ID No. 8 the amino acid sequence Gly-Ala-4Hyp-Gly-Lys-Asp-Gly-Val-Arg-Gly-Leu-Thr-Gly-Pro-Ile-Gly-Pro-4Hyp-Gly-Pro-Ala-Gly-Ala-4Hyp a 24-amino acid hydroxylated collagen peptide. SEQ ID No. 9 the amino acid sequence Gly-Ala-4Hyp-Gly-Lys-Asp-Gly-Val-Arg-Gly-Leu-Thr-Gly-Pro-Ile-Gly-Pro-4Hyp-Gly-Pro-Ala-Gly-Ala-Arg a 24-amino acid hydroxylated collagen peptide. SEQ ID No. 10 the amino acid sequence Gly-Ala-4Hyp-Gly-Pro-Pro-Gly-Pro-4Hyp-Gly-Ala-Arg-Gly-Gln-Ala-Gly-Val-Met-Gly-Phe-4Hyp-Gly-Pro-Lys a 24-amino acid hydroxylated collagen peptide. SEQ ID No. 11 the amino acid sequence Gly-Ala-Pro-Gly-Lys-Asp-Gly-Val-Arg-Gly-Leu an 11 amino acid non-hydroxylated collagen peptide. SEQ ID No. 12 the amino acid sequence Gly-Ala-Pro-Gly-Lys-Asp-Gly-Val-Arg-Gly-Leu-Thr-Gly-Pro-Ile-Gly-Pro-Pro-Gly-Pro-Ala-Gly-Ala a 23 amino acid non-hydroxylated collagen peptide. SEQ ID No. 13 the amino acid sequence Gly-Ala-Pro-Gly-Lys-Asp-Gly-Val-Arg-Gly-Leu-Thr-Gly-Pro-Ile-Gly-Pro-Pro-Gly-Pro-Ala-Gly-Ala-Pro-Gly-Asp-Lys-Gly-Glu-Ala-Gly-Pro-Ser-Gly-Pro-Ala-Gly-Pro-Thr-Gly-Ala-Arg-Gly-Ala-Pro-Gly-Asp-Arg-Gly-Glu-Pro-Gly-Pro-Pro-Gly-Pro-Ala-Gly a 58 amino acid non-hydroxylated collagen peptide. SEQ ID No. 14 the amino acid sequence Gly-Ala-Pro-Gly-Lys-Asp-Gly-Val-Arg-Gly-Leu-Thr-Gly-Pro-Ile-Gly-Pro-Pro-Gly-Pro-Ala-Gly-Ala-Pro-Gly-Asp-Lys-Gly-Glu-Ala-Gly-Pro-Ser-Gly-Pro-Ala-Gly-Pro-Thr-Gly-Ala-Arg-Gly-Ala-Pro-Gly-Asp-Arg-Gly-Glu-Pro-Gly-Pro-Pro-Gly-Pro-Ala-Gly-Phe-Ala-Gly-Pro-Pro-Gly-Ala-Asp-Gly-Gln-Pro-Gly-Ala-Lys-Gly-Glu-Pro-Gly-Asp-Ala-Gly-Ala-Lys-Gly-Asp-Ala-Gly-Pro-Pro-Gly-Pro-Ala a 90 amino acid non-hydroxylated collagen peptide. SEQ ID No. 15 the amino acid sequence Lys-Gly-Ala-Pro-Gly-Ala-Asp-Gly-Pro-Ala-Gly-Ala-Pro-Gly-Thr-Pro-Gly-Pro-Gln a 19 amino acid non-hydroxylated collagen peptide. SEQ ID No. 16 the amino acid sequence Gly-Pro-Pro-Gly-Pro-Ala-Gly-Glu-Lys-Gly-Ala-Pro-Gly-Ala-Asp-Gly-Pro-Ala-Gly-Ala-Pro-Gly-Thr-Pro-Gly-Pro-Gln-Gly-Ile-Ala-Gly-Gln-Arg-Gly-Val a 35 amino acid non-hydroxylated collagen peptide. SEQ ID No. 17 the amino acid sequence Gly-Ala-4Hyp-Gly-Lys-Asp-Gly-Val-Arg-Gly-Leu an 11 amino acid hydroxylated collagen peptide. SEQ ID No. 18 the amino acid sequence Gly-Ala-4Hyp-Gly-Lys-Asp-Gly-Val-Arg-Gly-Leu-Thr-Gly-Pro-Ile-Gly-Pro-4Hyp-Gly-Pro-Ala-Gly-Ala a 23-amino acid hydroxylated collagen peptide. SEQ ID No. 19 the amino acid sequence Gly-Ala-4Hyp-Gly-Lys-Asp-Gly-Val-Arg-Gly-Leu-Thr-Gly-Pro-Ile-Gly-Pro-4Hyp-Gly-Pro-Ala-Gly-Ala-4Hyp-Gly-Asp-Lys-Gly-Glu-Ala-Gly-Pro-Ser-Gly-Pro-Ala-Gly-Pro-Thr-Gly-Ala-Arg-Gly-Ala-4Hyp-Gly-Asp-Arg-Gly-Glu-4Hyp-Gly-Pro-4Hyp-Gly-Pro-Ala-Gly a 58 amino acid hydroxylated collagen peptide. SEQ ID No. 20 the amino acid sequence Gly-Ala-4Hyp-Gly-Lys-Asp-Gly-Val-Arg-Gly-Leu-Thr-Gly-Pro-Ile-Gly-Pro-4Hyp-Gly-Pro-Ala-Gly-Ala-4Hyp-Gly-Asp-Lys-Gly-Glu-Ala-Gly-Pro-Ser-Gly-Pro-Ala-Gly-Pro-Thr-Gly-Ala-Arg-Gly-Ala-4Hyp-Gly-Asp-Arg-Gly-Glu-4Hyp-Gly-Pro-4Hyp-Gly-Pro-Ala-Gly-Phe-Ala-Gly-Pro-4Hyp-Gly-Ala-Asp-Gly-Gln-4Hyp-Gly-Ala-Lys-Gly-Glu-Pro-Gly-Asp-Ala-Gly-Ala-Lys-Gly-Asp-Ala-Gly-Pro-Pro-Gly-Pro-Ala a 90 amino acid hydroxylated collagen peptide. SEQ ID No. 21 the amino acid sequence Lys-Gly-Ala-4Hyp-Gly-Ala-Asp-Gly-Pro-Ala-Gly-Ala-4Hyp-Gly-Thr-Pro-Gly-Pro-Gln a 19-amino acid hydroxylated collagen peptide. SEQ ID No. 22 the amino acid sequence Gly-Pro-4Hyp-Gly-Pro-Ala-Gly-Glu-Lys-Gly-Ala-4Hyp-Gly-Ala-Asp-Gly-Pro-Ala-Gly-Ala-4Hyp-Gly-Thr-Pro-Gly-Pro-Gln-Gly-Ile-Ala-Gly-Gln-Arg-Gly-Val a 35 amino acid hydroxylated collagen peptide. SEQ ID No. 23 the amino acid sequence Gly-Val-4Hyp-Gly-Lys-Tyr-Gly-Val-Arg-Gly-Leu-Thr-Gly-Pro-Ile-Gly-Pro-4Hyp-Gly-Pro-Leu-Gly-Ile-4Hyp-Gly-Trp-Lys-Gly-Phe-Val-Gly-Pro-Thr a 33 amino acid hydroxylated collagen peptide. SEQ ID No. 24 the amino acid sequence Gly-Pro-4Hyp-Gly-Pro-Val-Gly-Thr-Lys-Gly-Ile-4Hyp-Gly-Val-Tyr-Gly-Pro-Leu-Gly-Ile-4Hyp-Gly-Thr-Pro-Gly-Pro-Trp-Gly-Ile-Leu-Gly-Thr-Lys-Arg-Gly-Val a 36 amino acid hydroxylated collagen peptide. SEQ ID No. 25 the amino acid sequence Gly-Ala-4Hyp-Gly-Lys-Asp-Gly-Val-Arg-Gly-Leu-Thr-Gly-4Hyp-Ile-Gly-4Hyp-4Hyp-Gly-4Hyp-Ala-Gly-Ala-4Hyp-Gly-Asp-Lys-Gly-Glu-Ala-Gly-4Hyp-Ser a 33 amino acid hydroxylated collagen peptide. SEQ ID No. 26 the amino acid sequence His-His-His-His-His-His-Gly-Ala-4Hyp-Gly-Lys-Asp-Gly-Val-Arg-Gly-Leu-Thr-Gly-Pro-Ile-Gly-Pro-4Hyp-Gly-Pro-Ala-Gly-Ala-4Hyp-Gly-Asp-Lys-Gly-Glu-Ala-Gly-Pro-Ser a 39 amino acid hydroxylated collagen peptide. SEQ ID No. 27 the amino acid sequence Gly-Leu-4Hyp-Gly-Lys-Met-Gly-Val-Phe-Gly-Leu-Thr-Gly-Pro-Ile-Gly-Pro-4Hyp-Gly-Pro-Trp-Gly-Val-4Hyp-Gly-His-Lys-Gly-Tyr-Leu-Gly-Pro-Thr a 33 amino acid hydroxylated collagen peptide. SEQ ID No. 28 the amino acid sequence Gly-Leu-4Hyp-Gly-Lys-Tyr-Gly-Val-His-Gly-Leu-Thr-Gly-Pro-Leu-Gly-Pro-4Hyp-Gly-Pro-Met-Gly-Ile-4Hyp-Gly-Trp-Lys-Gly-Phe-Val-Gly-Pro-Thr a 33 amino acid hydroxylated collagen peptide. SEQ ID No. 29 the amino acid sequence Gly-Pro-4Hyp-Gly-Pro-Leu-Gly-Met-Lys-Gly-Leu-4Hyp-Gly-Val-Trp-Gly-Pro-Phe-Gly-Leu-4Hyp-Gly-Thr-Pro-Gly-Pro-His-Gly-Ile-Thr-Gly-Tyr-Lys-Gly-Val a 35 amino acid hydroxylated collagen peptide. SEQ ID No. 30 the amino acid sequence Gly-Pro-4Hyp-Gly-Pro-Val-Gly-Thr-Lys-Gly-Leu-4Hyp-Gly-Phe-Tyr-Gly-Pro-Leu-Gly-Ile-4Hyp-Gly-His-Pro-Gly-Pro-Trp-Gly-Met-Leu-Gly-Thr-Lys-Gly-Val a 35 amino acid hydroxylated collagen peptide.

[0179] It shows Figure 1AThe stimulation of collagen synthesis in human dermal fibroblasts by the collagen peptides of SEQ ID No. 2, SEQ ID No. 7, SEQ ID No. 8, SEQ ID No. 9, and SEQ ID No. 10 according to the invention compared to a mixture of collagen peptides obtained from natural sources with a comparable average molecular weight (Verisol) and compared to two different gelatin controls (160 Bloom gelatin from pork rinds (Gelatin 160), 260 Bloom gelatin from beef split skin (Gelatin 260)) according to Example 6. The factor for the quantitative determination of collagen synthesis compared to an untreated sample is shown. The error bars indicate the standard error (SEM) in each case. Figure 1BThe stimulation of collagen synthesis in human chondrocytes by the collagen peptides of SEQ ID No. 2, SEQ ID No. 7, SEQ ID No. 8, SEQ ID No. 9, and SEQ ID No. 10 according to the invention compared to a mixture of collagen peptides obtained from natural sources with a comparable average molecular weight (Fortigel) according to Example 6. The factor for the quantitative determination of collagen synthesis compared to an untreated sample is shown. The error bars indicate the standard error (SEM). Figure 2AThe stimulation of proteoglycan synthesis in human dermal fibroblasts by the collagen peptides of SEQ ID No. 2, SEQ ID No. 7, SEQ ID No. 8, SEQ ID No. 9, and SEQ ID No. 10 according to the invention compared to a mixture of collagen peptides obtained from natural sources with a comparable average molecular weight (Verisol) and compared to two different gelatin controls (160 Bloom gelatin from pork rinds (Gelatin 160), 260 Bloom gelatin from beef split skin (Gelatin 260)) according to Example 6. The factor for the quantitative determination of proteoglycan synthesis compared to an untreated sample is shown. The error bars indicate the standard error (SEM) in each case. Figure 2BThe stimulation of the synthesis of proteoglycans of human chondrocytes by the collagen peptides according to the invention of SEQ ID No. 2, SEQ ID No. 7, SEQ ID No. 8, SEQ ID No. 9, SEQ ID No. 10 compared to a mixture of collagen peptides obtained from natural sources with a comparable average molecular weight (Fortigel) according to Example 6. The factor for the quantitative determination of collagen synthesis compared to an untreated sample is shown. The error bars indicate the standard error (SEM). Figure 3The stimulation of collagen synthesis in human chondrocytes by the inventive collagen peptides of SEQ ID No. 11 to SEQ ID No. 26 compared to a mixture of collagen peptides obtained from natural sources with a comparable average molecular weight (Fortigel). The factor for the quantitative determination of collagen synthesis compared to an untreated sample is shown. The error bars indicate the standard error (SEM). Figure 4 The stimulation of collagen synthesis in human dermal fibroblasts by the collagen peptides of the invention, SEQ ID No. 11 to SEQ ID No. 26, compared to a mixture of collagen peptides obtained from natural sources with a comparable average molecular weight (Verisol). The factor for the quantitative determination of collagen synthesis compared to an untreated sample is shown. The error bars indicate the standard error (SEM). Figure 5The stimulation of proteoglycan synthesis in human chondrocytes by the collagen peptides of the invention, SEQ ID No. 11 to SEQ ID No. 24, compared to a mixture of collagen peptides obtained from natural sources with a comparable average molecular weight (Fortigel). The factor for the quantitative determination of collagen synthesis compared to an untreated sample is shown. The error bars indicate the standard error (SEM). Figure 6 The stimulation of proteoglycan synthesis in human dermal fibroblasts by the collagen peptides of SEQ ID No. 11 to SEQ ID No. 26 according to the invention compared to a mixture of collagen peptides obtained from natural sources with a comparable average molecular weight (Verisol). The factor for the quantitative determination of collagen synthesis compared to an untreated sample is shown. The error bars indicate the standard error (SEM). Figure 7AThe stimulation of collagen synthesis in human dermal fibroblasts by the inventive collagen peptides of SEQ ID No. 27 to SEQ ID No. 30 compared to a mixture of collagen peptides obtained from natural sources with a comparable average molecular weight (Verisol). The factor for the quantitative determination of collagen synthesis compared to an untreated sample is shown. The error bars indicate the standard error (SEM). Figure 7B The stimulation of proteoglycan synthesis in human dermal fibroblasts by the collagen peptides of SEQ ID No. 27 to SEQ ID No. 30 according to the invention compared to a mixture of collagen peptides obtained from natural sources with a comparable average molecular weight (Verisol). The factor for the quantitative determination of collagen synthesis compared to an untreated sample is shown. The error bars indicate the standard error (SEM). Examples: Example 1 - Solid-phase synthesis

[0180] Collagen peptides according to the invention of the amino acid sequences SEQ ID No. 1 to SEQ ID No. 10 and SEQ ID No. 27 to SEQ ID No. 30 were obtained by solid-phase synthesis (Merrifield synthesis) on a polystyrene resin. Example 2 - recombinant production

[0181] Further collagen peptides, in particular the collagen peptides of the amino acid sequences SEQ ID No. 1 to SEQ ID No. 10, were additionally produced by recombinant expression in Pichia pastoris won. Example 3 - Bone health:

[0182] To analyze the biological efficacy of the collagen peptide according to the invention with regard to maintaining bone health and the prevention and treatment of bone diseases, its stimulating effect on the synthesis of extracellular matrix proteins and enzymes that play a role in matrix formation and mineralization by osteoblasts is investigated in vitro. This is done by determining the expression of the corresponding mRNA using real-time PCR and a semi-quantitative evaluation (relative to a control without collagen peptide).

[0183] For this purpose, human osteoblasts are first isolated from knee joints by incubating bone material under vigorous agitation at 37°C for 1 h in Hanks' solution supplemented with 7 mg / ml hyaluronidase types I and III-S and 5 mg / ml pronase. The digestion is then continued at 37°C for 3–5 h in Hanks' solution supplemented with 16 mg / ml collagenase type CLS IV. After enzymatic digestion, the resulting primary osteoblasts are cultured in Ham's F12 medium supplemented with 10% fetal calf serum, 20 U / ml penicillin-streptomycin, 50 µg / ml partricin, 0.05 mg / ml ascorbic acid, and 0.15 mg / ml glutamine. Alternatively, primary osteoblasts (item no. C-12760; 2019) can also be purchased from PromoCell GmbH, Heidelberg, Germany, for testing their biological efficacy. The cells are then cultured in Ham's F12 medium supplemented with 10% fetal calf serum, 20 U / ml penicillin-streptomycin, 50 µg / ml partricin, and 0.15 mg / ml glutamine.

[0184] To investigate biological efficacy, monolayer cell cultures of isolated human osteoblasts are incubated for 24 hours in medium supplemented with 0.5 mg / ml of the respective collagen peptide. A control is incubated in medium without peptide. Subsequently, the respective mRNA expression is determined. Example 4 - Skin health:

[0185] The stimulation of collagen (type I) synthesis and the proteoglycans biglycan and versican is investigated in vitro in human dermal fibroblasts (skin cells). For this purpose, the cells are incubated for 24 hours with 0.5 mg / ml of a low-molecular-weight collagen peptide or the collagen peptide of the invention. The expression of collagen RNA, biglycan RNA, and versican RNA is then determined using real-time PCR and evaluated semiquantitatively (relative to a control without peptide). Example 5 - Cartilage health:

[0186] For cell culture, porcine or human chondrocytes are isolated from cartilage tissue in a conventional manner and seeded onto culture plates at a density of approximately 350,000 cells / cm². The culture medium used is Ham's F12 medium containing 10% fetal calf serum, 10 µg / ml gentamicin, and 5 µg / ml amphotericin B. Alternatively, 10 µg / ml penicillin-streptomycin can be used instead of 10 µg / ml gentamicin. Cultivation was carried out at 37°C in an oxygen-reduced atmosphere (5% O², 5% CO², and 90% N²).

[0187] Determination of collagen biosynthesis: The quantification of collagen synthesized by chondrocytes (mainly type II) is carried out by radioactive labeling with 14< C-proline, which is incorporated into the collagen.

[0188] Radioactive 14<C-proline is first added to the culture medium, and the chondrocytes are cultured under these conditions until the time of determination. To distinguish incorporated from unincorporated 14<C-proline during detection, the isotope-containing culture medium is then replaced with pure culture medium for a period of 3 days. The culture medium is then discarded, and the adherent cell layer is treated with distilled water to disrupt the cell membranes through osmotic stress and release cytosolic, unbound 14<C-proline. The cell debris is pelleted with the synthesized extracellular matrix by centrifugation. The pellet is resuspended in fresh distilled water and mixed with a xylene scintillation cocktail. The amount of synthesized collagen can then be quantified by detecting the 14<C-proline with a beta counter.

[0189] Alternatively, quantification can be performed using the Sircol Collagen Assay Kit (Article No. 054S5000, 2019, tebu-bio, Offenbach, Germany, or Biocolor Ltd., UK) according to the manufacturer's instructions (see Example 6).

[0190] Determination of proteoglycan biosynthesis: The quantification of the proteoglycans synthesized by the chondrocytes is carried out by Alcian blue staining and photometric determination of the glycosaminoglycans (GAGs), which are components of the proteoglycans.

[0191] To determine the GAG content in the cell culture, the culture medium is first discarded, and the adherent cell layer is rinsed with PBS buffer (pH 7). The cells are then fixed at 4°C for 2 hours in a 10% formaldehyde solution in PBS. After the formaldehyde is removed, the Alcian Blue staining reagent (5% Alcian Blue in 3% acetic acid) is applied to the cell layer and incubated overnight at 4°C. Unbound Alcian Blue is discarded and washed out by gently rinsing three to four times with PBS. The GAG complexes are released from the cell layer by adding acidic guanidine solution (8 mol / L). The amount of glycosaminoglycans can then be quantified photometrically at a wavelength of 620 nm.

[0192] Alternatively, quantification can be performed using the Blyscan Glycosaminoglycan Assay Kit (Article No. 054B3000, 2019, tebu-bio, Offenbach, Germany, or Biocolor Ltd., UK) according to the manufacturer's instructions (see Example 6). Example 6 - Influence of synthetic collagen peptides according to the invention on the biosynthesis of matrix proteins Cell culture:

[0193] The human cells used were obtained from tebu-bio GmbH, Offenbach, Germany.

[0194] The chondrocytes (cat. no. 402-05a) or dermal fibroblasts (cat. no. 106-05a) were first seeded into 12-well culture plates and cultivated at 37°C, 5% CO2 in Ham's F12 medium to which 10% fetal calf serum, 20 U / ml penicillin-streptomycin and 50 µg / ml ascorbic acid were added. Every other day the culture medium was replaced with new culture medium until a cell confluence of 80% was reached. To investigate the influence of the collagen peptides according to the invention of SEQ ID No. 2 (non-hydroxylated), SEQ ID No. 7 (hydroxylated), SEQ ID No. 8 (hydroxylated), SEQ ID No. 9 (hydroxylated), SEQ ID No. 10 (hydroxylated), SEQ ID No. SEQ ID No. 11 (non-hydroxylated), SEQ ID No. 12 (non-hydroxylated), SEQ ID No. 13 (non-hydroxylated), SEQ ID No. 14 (non-hydroxylated), SEQ ID No. 15 (non-hydroxylated), SEQ ID No. 16 (non-hydroxylated), SEQ ID No. 17 (hydroxylated), SEQ ID No. 18 (hydroxylated), SEQ ID No. 19 (hydroxylated), SEQ ID No. 20 (hydroxylated), SEQ ID No.To determine the effect of SEQ ID No. 21 (hydroxylated), SEQ ID No. 22 (hydroxylated), SEQ ID No. 23 (hydroxylated), SEQ ID No. 24 (hydroxylated), SEQ ID No. 25 (hydroxylated), SEQ ID No. 26 (hydroxylated), SEQ ID No. 27 (hydroxylated), SEQ ID No. 28 (hydroxylated), SEQ ID No. 29 (hydroxylated) and SEQ ID No. 30 (hydroxylated) on the biosynthesis of matrix proteins, the cell culture medium was subsequently replaced by a special stimulation medium to which 0.5 mg / ml of the specific collagen peptides had been added. Collagen assay:

[0195] To investigate collagen metabolism, the amount of newly synthesized collagen was determined after three weeks of cell stimulation with 0.5 mg / ml BCP. The synthesized collagen was isolated using the Sircol assay (item no. 054S5000, 2019, tebu-bio, Offenbach, Germany, or Biocolor Ltd., UK) according to the manufacturer's instructions. Briefly, the culture medium was first discarded, and the adherent cell layers were digested with 0.1 mg of pepsin solution in 0.5 M acetic acid at 4 °C overnight. The cell suspensions were neutralized by adding 100 µl of an acid-neutralizing reagent. The synthesized collagen was then separated by adding 200 µl of an isolation and concentration solution with vigorous shaking at 4 °C overnight. After centrifugation (12,000 rpm, 10 min) and discarding the supernatant, the isolated collagen was resuspended in 1 ml of Sircol dye solution.After 30 minutes of shaking and another centrifugation, the collagen pellet was overlaid with 750 µl of cold acidic solid wash reagent. After further centrifugation, the supernatant was discarded, and the enriched collagen was resuspended in 250 µl of alkaline solution. 200 µl of each sample solution was used for photometric quantification of the synthesized collagen. The absorbance was measured at a wavelength of 492 nm. The amount of synthesized collagen was determined based on standardized collagen solutions.

[0196] In comparison to untreated controls, an increased collagen synthesis was found by dermal fibroblasts which had been treated with the hydroxylated collagen peptides according to the invention of SEQ ID No. 7 (42% increase), SEQ ID No. 8 (37% increase), SEQ ID No. 9 (38% increase), SEQ ID No. 10 (35% increase), SEQ ID No. 17 (42% increase), SEQ ID No. 18 (40% increase), SEQ ID No. 19 (40% increase), SEQ ID No. 20 (37% increase), SEQ ID No. 21 (36% increase), SEQ ID No. 22 (43% increase), SEQ ID No. 23 (39% increase), SEQ ID No. 24 (36% increase), SEQ ID No. 25 (27% increase), SEQ ID No. 26 (12% increase), SEQ ID No. 27 (27% increase), SEQ ID No. 28 (22% increase), SEQ ID No. 29 (21% increase) and SEQ ID No. 30 (21% increase) were incubated ( Fig. 1A ; Fig. 4 ; Fig. 7A). Furthermore, it was shown that the incubation of dermal fibroblasts with non-hydroxylated collagen peptides according to the invention causes at least a comparable stimulation of collagen synthesis as the incubation of the cells with a mixture of hydroxylated collagen peptides obtained from natural sources with a comparable average molecular weight (Verisol) (SEQ ID No. 2 (22% increase), SEQ ID No. 11 (19% increase), SEQ ID No. 12 (22% increase), SEQ ID No. 13 (20% increase), SEQ ID No. 14 (22% increase), SEQ ID No. 15 (23% increase) and SEQ ID No. 16 (22% increase)). All collagen peptides according to the invention investigated showed a significant increase in collagen synthesis compared to gelatin controls (Gelatin 160, Gelatin 260).

[0197] Increased collagen synthesis was also observed for chondrocytes incubated with the collagen peptides according to the invention compared to untreated controls. Incubation of chondrocytes with hydroxylated collagen peptides led to an increase in collagen synthesis of 28% (SEQ ID No. 7), 24% (SEQ ID Nos. 8, 9), 20% (SEQ ID No. 10), 33% (SEQ ID No. 17), 32% (SEQ ID No. 18), 37% (SEQ ID No. 19), 30% (SEQ ID No. 20), 55% (SEQ ID No. 21), 46% (SEQ ID No. 22), 19% (SEQ ID Nos. 23, 25), 26% (SEQ ID No. 24), and 12% (SEQ ID No. 26). The incubation of chondrocytes with non-hydroxylated collagen peptides according to the invention resulted in at least a comparable stimulation of collagen synthesis as the incubation of cells with a mixture of hydroxylated collagen peptides obtained from natural sources with a comparable average molecular weight (Fortigel) ( Fig. 1B ; Fig. 3). Incubation of chondrocytes with non-hydroxylated collagen peptides led to an increase in collagen synthesis by 16% (SEQ ID No. 2, 11), 24% (SEQ ID No. 12), 15% (SEQ ID No. 13), 28% (SEQ ID No. 14) and 31% (SEQ ID Nos. 15, 16). Proteoglycan assay:

[0198] The Blyscan Glycosaminoglycan Assay (Article No. 054B3000, 2019, tebu-bio, Offenbach, Germany, or Biocolor Ltd., UK) was used to determine proteoglycan biosynthesis after two weeks of stimulation of dermal fibroblasts. According to the manufacturer's instructions, the cell layers were overlaid with 1 ml of papain extraction solution after discarding the culture medium and incubated for three hours at 65°C with vigorous shaking. The cell suspensions were then centrifuged (10,000 g, 10 min), and the supernatants were collected. After adding 1 ml of Blyscan dye solution and shaking (30 min), the supernatants were centrifuged again (12,000 rpm, 10 min) and then discarded. The isolated proteoglycan pellets were resuspended in 500 µl of dissociation solution.A photometric determination of the synthesized proteoglycan in 200 µl sample solution was carried out at a wavelength of 656 nm in comparison to untreated control experiments.

[0199] As from Fig. 2A , 6 and 7BAs can be seen, the synthesis of proteoglycans by dermal fibroblasts was significantly increased when the fibroblasts were incubated with collagen peptides according to the invention compared to untreated controls. Incubation of the cells with hydroxylated collagen peptides led to an increase in the synthesis of proteoglycans by 41% (SEQ ID No. 7), 23% (SEQ ID No. 8, 25), 27% (SEQ ID No. 9, 21), 25% (SEQ ID No. 10), 24% (SEQ ID No. 17), 31% (SEQ ID No. 18), 29% (SEQ ID Nos. 19, 20), 28% (SEQ ID No. 22), 22% (SEQ ID No. 23, 28, 30), 26% (SEQ ID No. 24, 27), 6% (SEQ ID No. 26) and 21% (SEQ ID No. 29).It was also shown that incubation of dermal fibroblasts with non-hydroxylated collagen peptides according to the invention results in at least a comparable stimulation of proteoglycan synthesis as incubation of the cells with a mixture of hydroxylated collagen peptides obtained from natural sources with a comparable average molecular weight (Verisol). Incubation of dermal fibroblasts with non-hydroxylated collagen peptides led to an increase in collagen synthesis of 18% (SEQ ID Nos. 2, 11, 15, 16), 19% (SEQ ID No. 12), 16% (SEQ ID No. 13), and 14% (SEQ ID No. 14). A significant increase in proteoglycan synthesis was observed for all collagen peptides according to the invention tested compared to gelatin controls (Gelatin 160, Gelatin 260).

[0200] Chondrocytes incubated with the collagen peptides according to the invention also show an increased synthesis of proteoglycans compared to untreated controls ( Fig. 2B and 5 ). Incubation of chondrocytes with hydroxylated collagen peptides led to an increase in collagen synthesis by 30% (SEQ ID No. 7), 20% (SEQ ID No. 8), 19% (SEQ ID No. 9), 29% (SEQ ID No. 10), 31% (SEQ ID No. 17), 25% (SEQ ID No. 18), 18% (SEQ ID No. 19), 15% (SEQ ID No. 20, 24), 22% (SEQ ID No. 21), 26% (SEQ ID No. 22, 24) and 11% (SEQ ID No. 23). The incubation of chondrocytes with non-hydroxylated collagen peptides according to the invention resulted in at least a comparable stimulation of the synthesis of proteoglycans as the incubation of cells with a mixture of hydroxylated collagen peptides obtained from natural sources with a comparable average molecular weight (Fortigel) ( Fig. 5 ; Fig. 2B). Incubation of chondrocytes with non-hydroxylated collagen peptides led to an increase in the synthesis of proteoglycans by 22% (SEQ ID No. 2), 28% (SEQ ID No. 11), 23% (SEQ ID No. 12), 9% (SEQ ID No. 13), 14% (SEQ ID No. 14), 6% (SEQ ID No. 15) and 18% (SEQ ID No. 16).

Claims

1. A synthetic or recombinant collagen peptide for use in a method for the prevention and / or treatment of bone diseases, cartilage diseases, degenerative joint diseases, diseases of the tendons or ligaments and / or treatment of skin diseases, wherein the collagen peptide has a molecular weight in a region of 0.18 to 5.0 kDa and is able to stimulate the synthesis of extracellular matrix proteins, in particular collagen, in connective tissue cells, in particular in osteoblasts, chondrocytes and / or fibroblasts, and wherein the collagen peptide has an amino acid sequence selected from the group consisting of SEQ ID No. 1 to 30.

2. The collagen peptide for use of claim 1, wherein the collagen peptide has the amino acid sequence SEQ ID No. 1 or 15.

3. The collagen peptide for use of claim 1 or 2, wherein the collagen peptide has the amino acid sequence SEQ ID No. 1, in which at least one proline residue is hydroxylated, in particular wherein the collagen peptide has the amino acid sequence SEQ ID No. 6.

4. The collagen peptide for use of one of the preceding claims 1 to 3, wherein the collagen peptide has an amino acid sequence selected from the group consisting of SEQ ID No. 1 to 4, 6 to 9, 12 to 14 and 18 to 20.

5. The collagen peptide for use of claim 1 or 2, wherein the collagen peptide has no amino acid modification, in particular no hydroxylation.

6. The collagen peptide for use of one of claims 1 to 4, wherein the collagen peptide has hydroxylated proline, hydroxylated lysine or hydroxylated proline and lysine.

7. The collagen peptide for use of one of claims 1 to 4 or 6, wherein the collagen peptide is glycosylated at at least one hydroxylated lysine.

8. A pharmaceutical composition comprising a synthetic or recombinant collagen peptide with a molecular weight in a region of 0.18 to 5.0 kDa, which is able to stimulate the synthesis of extracellular matrix proteins, in particular collagen, in connective tissue cells, in particular in osteoblasts, chondrocytes and / or fibroblasts, and at least one pharmaceutically acceptable additive, wherein the collagen peptide has an amino acid sequence selected from the group consisting of SEQ ID No. 1 to 30 and wherein no further collagen peptides are present in the composition.

9. A food supplement, foodstuff or luxury foodstuff comprising a synthetic or recombinant collagen peptide with a molecular weight in a region of 0.18 to 5.0 kDa, which is able to stimulate the synthesis of extracellular matrix proteins, in particular collagen, in connective tissue cells, in particular in osteoblasts, chondrocytes and / or fibroblasts, and at least one food-acceptable additive, wherein the collagen peptide has an amino acid sequence selected from the group consisting of SEQ ID No. 1 to 30 and wherein no further collagen peptides are present in the food supplement, foodstuff or luxury foodstuff.

10. A cosmetic product comprising a synthetic or recombinant collagen peptide with a molecular weight in a region of 0.18 to 5.0 kDa, which is able to stimulate the synthesis of extracellular matrix proteins, in particular collagen, in connective tissue cells, in particular in osteoblasts, chondrocytes and / or fibroblasts, optionally together with at least one skin-compatible additive, wherein the collagen peptide has an amino acid sequence selected from the group consisting of SEQ ID No. 1 to 30 and wherein no further collagen peptides are present in the product.

11. A non-therapeutic method for optically improving the skin, in particular for reducing the formation of wrinkles, improving skin elasticity, increasing skin tone, increasing the moisture content of the skin, reducing cellulite and / or reducing stretch marks, in particular pregnancy stretch marks, accelerating nail growth and / or reducing nail brittleness, optically improving the hair, in particular improving hair quality, decreasing split ends and / or reducing or delaying hair loss, wherein at least one synthetic or recombinant collagen peptide with a molecular weight in a region from 0.18 to 5.0 kDa is administered to the human or animal body, which is able to stimulate the synthesis of extracellular matrix proteins, in particular collagen, in connective tissue cells, in particular in osteoblasts, chondrocytes and / or fibroblasts, and wherein the collagen peptide has an amino acid sequence selected from the group consisting of SEQ ID No. 1 to 30.